Philip Petra
Emeritus Professor of Biochemistry

 

CURRICULUM VITAE

PHILIP HUE PETRA

(Revised – 2000, 2010, 2023)

Personal Data

Birthday: May 12, 1937
Birthplace: Paris, France
Citizenship: U.S., 1962
Marital Status: Married, Dolores
Children: Philippe
Anita

Education

B.S. Department of Chemistry
Tulane University
New Orleans, LA		 1955-60
M.S. Department of Biochemistry
Tulane University
New Orleans, LA
Dr. Elliot Shaw, thesis advisor
Thesis: A kinetic analysis of the catalyzed hydrolysis of α-N-benzoyl-l-citrulline methyl ester by papain		 1960-62
Ph.D. Department of Biochemistry
Tulane University
New Orleans, LA
Dr Gunther Schoellman and Dr. Elliott N. Shaw, thesis advisors
Thesis: Isolation and characterization of the alkylated histidine residue from trypsin inhibited by a specific reagent, TLCK		 1962-66
Postdoctoral Research Professor Hans Neurath
Department of Biochemistry
University of Washington Seattle, WA
Research: Structure-function relationship of Carboxypeptidase A		 1966-70

Academic Appointments

Position Departments Institution Dates
Acting Assistant Professor Department of Biochemistry University of Washington, Seattle, WA 1968-70
Assistant Professor Department of Obstetrics/Gynecology (joint with Biochemistry 1970-74; adjunct with Biochemistry 1974-75) University of Washington, Seattle, WA 1970-75
Assistant Professor with tenure Departments of Ob/Gyn and Biochemistry University of Washington, Seattle, WA 1975-78
Associate Professor Departments of Ob/Gyn and Biochemistry University of Washington, Seattle, WA 1978-84
Professor Departments of Ob/Gyn and Biochemistry University of Washington, Seattle, WA 1984-93
Professor Department of Biochemistry University of Washington, Seattle, WA 1993-2006
Professor Emeritus of Biochemistry – University of Washington, Seattle, WA 2006 –

Societies

  • The American Society for Biochemistry and Molecular Biology
  • The Protein Society
  • The Endocrine Society
  • The American Association for the Advancement of Science

Honors and Invitations

  • Exchange Scientist of the National Cancer Institute with Institut National de la Sante et Recherche Medicale, Paris, France, 1977.
  • Speaker, International Congress on Hormonal Steroids, New Delhi, India, 1978.
  • Speaker, International Congress on Hormonal Steroids, Jerusalem, Israel, 1982.
  • Symposium and speaker, Society for Gynecological Investigation, Washington, D.C., 1983.
  • Speaker, 7th International Symposium of the Journal of Steroid Biochemistry, Seefeld, Austria, 1985.
  • Visiting Professor with Dr. Etienne-Emile Baulieu, Institut National de la Sante et de la Recherche Medicale (INSERM) Paris, France, Sept 1985 – Sept 1986.
  • Seminar speaker, University of Geneva, Geneva, Switzerland, February 1986.
  • Seminar speaker, CNRS, Caen, France, March 1986.
  • Scientific Organizing Committee and speaker, First International Symposium on Steroid Hormone Binding Proteins, Lyon, France, April 1986.
  • Seminar speaker, INSERM U34, Lyon, France, May 1986.
  • Scientific Organizing Committee and speaker, 2nd International Congress on Steroid-Protein Interaction, Torino, Italy, September 1987.
  • Scientific Organizing Committee and speaker, 3rd International Congress on Steroid Binding Proteins, The Hague, The Netherlands, September 1990.

Teaching Responsibilities

1. Training responsibilities on NIH Ob/Gyn Departmental Training Grant. 1971 – 1978
2. Biochemistry 406 – General principles of Biochemistry
Class size – 250-500		 1978 – 2006
3. Biochemistry 426 – Basic Techniques in Biochemistry.
Class size – 40 students.		 1994 – 2000
4. Biochemistry 499 – Undergraduate Research. 1978 – 2000
5. Hu-BIO 514 1996 – 1997
6. Biochemistry 540 – Literature Review for biochemistry graduate students.
7. Biochemistry 581 – Laboratory rotation for biochemistry graduate students.

Research Funding: Principal Investigator

1. Graduate School Research Fund (Univ. of Wash.)
Title: Steroid Binding Proteins.
December 1971 – December 1972		 $2,935
2. American Cancer Society Institutional Grant.
Title: Biochemical studies on estradiol induction of metabolism in the uterus.
February 1973 – January 1974		 $2,400
3. NIH. The National Institute of Child Health and Human Development.
Title: Structure-Function of Steroid-binding Proteins
May 1972 – April 1975		 $94,400
4. American Cancer Society Institutional Grant.
Title: A filter assay for estrogen receptors in human mammary tumors.
December 1974 – December 1975.		 $1,200
5. NIH. The National Institute of Child Health and Human Development.
Title: Structure-Function of Steroid-binding Proteins.
June 1975 – May 1979.		 $124,114
6. American Cancer Society.
Title: Study of breast neoplasia with advanced receptor methodology.
July 1976 – July 1978.		 $71,085
7. Biomedical Research Support Grant (Univ. of Wash.).
Title: Influence of Steroid-binding plasma proteins on the growth of estrogen-dependent breast cancer.
July 1978 – March 1979.		 $4,356
8. NIH. The National Institute of Child Health and Human Development.
Title: Characterization of the plasma sex steroid-binding protein.
April 1981 – March 1988.		 $568,243
9. NIH. The National Institute of Child Health and Human Development
Title: Characterization of the plasma sex steroid-binding protein.
April 1988 – March 1993.		 $620,000
10. ROYALTY RESEARCH FUNDS, University of Washington
Title: Cloning of the Sex Steroid-Binding Protein.
January 1994 – May 1995.		 $34,230
11. NIH. The National Institute of Child Health and Human Development.
Title: Structure of plasma sex steroid-binding protein.
April 1995 – May 2001.		 $400,000

 

Research Activity

My interest in structure-function relationships in proteins began during my graduate work at Tulane University in 1960 under the guidance of Drs Elliott Shaw and Gunther Schoellman who had originated the method of affinity labelling of proteins. That chemical approach consists of designing small reactive molecules, resembling enzyme substrates, in the hope that they would bind and react with amino acid side chains in the active site thus identifying them as possible catalytic groups. Using this approach, they were able to identify a histidine residue in the active site of chymotrypsin, later identified as His-47, which functions in the hydrolytic action of the enzyme. At the time, only a few sequences of proteins were known (insulin, ribonuclease, and lysozyme) and both chymotrypsin and trypsin had not yet been determined. For my PhD thesis, I was given the task of characterizing the chemical attack of the analogous histidine residue in trypsin by the affinity label, L-1-chloro3-tosylamido-7-amino-2-heptanone (TLCK), a chloro-methyl ketone which Shaw and co-workers had shown to inhibit trypsin in a 1:1 mole ratio. I accomplished this task and found that the reactive group of the affinity label had attacked position 3 of the imidazole ring of a histidine residue (1), later identified as His-57 when Walsh et al. solved the amino acid sequence of trypsin.

After completion of my thesis, I moved to Seattle as a postdoctoral fellow with Hans Neurath. My first task was to solve the purification of bovine carboxypeptidase (CP-A) which had been elusive (3,5). I then turned my attention to the active center of the enzyme, the amino acid sequence of which had now been solved by Bradshaw, Walsh, and Neurath. The 3-D structure, solved by Lipscomb at Harvard, revealed the presence of two glutamic acid residues in the active center (Glu-72 and Glu-270), one possibly serving as catalytic group. I was given the task of resolving that question. N-ethyl-5-phenylisoxazolium-3’-sulfonate (reagent K) was chosen as a carboxyl group-specific reagent because this reagent incorporates some of the structural features of small substrates of CP-A and thus could serve as an affinity label. R. B. Woodward had used it for organic synthesis, and the Tulane Shaw group had used a similar reagent for the modification of carboxyl groups in proteins. Reagent K was found to inhibit CP-A in the absence of competitive inhibitors but not in their presence. The label attached to a carboxyl group in the enzyme active site, presumably a glutamic acid, as a mixed anhydride. I chose to stabilize it by nucleophilic displacement with 14C-methoxamine converting it to a stable amide linkage. The labelled residue was identified as Glu-270 by sequencing radioactive peptides (6, 7). The data identified Glu-270 as the catalytic group in enzymatic hydrolysis.

After completing other projects in the Neurath group, I started my faculty appointment at the University of Washington in 1970. I was offered an assistant professorship by Walter Hermann, chairman of the department of Ob/Gyn, and by Hans Neurath, chairman of the department of Biochemistry. The appointment was joint with teaching responsibilities in biochemistry, and research responsibilities in Ob/Gyn. I directed my attention to steroid-binding proteins which were implicated in steroid hormone action. These are present in plasma and target cells and bind androgens (testosterone T, and dihydrotestosterone DHT), estrogens (E2), and glucocorticoids at high affinity. Dr. Hermann et al. had discovered an estrogen receptor in brain tissue, so there was already experience in this field in the Ob/Gyn department. At the time, none of these proteins had been characterized, sequenced, or cloned. The major problem was their low concentrations in both cells and plasma. I decided to focus on the human plasma protein called SBP (Sex Steroid Binding protein), also called SHBG (Sex Hormone Binding Globulin) which had been discovered in Etienne Baulieu’s group in Paris in the 1960s. I wrote my first NIH grant in 1972 and was able to secure funding. Because hSBP increases significantly in pregnancy, we obtained serum left over in the Ob/Gyn Delivery Rooms. The low plasma levels of human SBP (hSBP) required development of an efficient purification method. A reactive DHT derivative still binding to hSBP was needed to construct an affinity adsorbent. Dr. Niels Anderson of the chemistry Department at the University of Washington helped us synthesize such a derivative (22). Non-human primate plasma was obtained from the Oregon Primate Center in collaboration with Miles Novy and Frank Stanczyk. This led to the purification and characterization of hSBP (16), macaque nemestrina SBP (nSBP) (30), macaque mulatta SBP (rhSBP) (30), baboon SBP (bSBP) (39), and rSBP (rabbit SBP) (17) as well as determination of the amino acid sequences of hSBP and rSBPs (41,50). A radioactive affinity label, bromomethyl ketone derivative of DHT, [14C]DHTBr, was synthesized and led to the identification of Lys-134 in the steroid binding site (53). The cDNA-derived amino acid sequence of the steroid binding domain of the human androgen receptor had just been determined; it appeared to bear no similarity to hSBP. This held true for other sex steroid receptors suggesting that steroid-binding proteins belong to at least two different gene families. Moreover, the gene products of these families bind their respective steroids with similar Kds (0.2nM-1nM) suggesting convergent molecular evolution. All SBPs are homodimeric glycoproteins with a single steroid binding site. The hSBP monomer is a 373-residue polypeptide containing three carbohydrate chains, one O-linked to Th-7, and the other two N-linked to Asn-351 and Asn-367, and two disulfide bonds Cys164-Cys188 and Cys333-Cys361. Rabbit SBP is shorter by 6 residues at the N-terminus, has two N-linked carbohydrate side chains at homologous positions, but lacks the O-linked carbohydrate chain (50).  Human SBP and rSBP are 80% homologous, both sequences contain one tyrosine residue at homologous positions (Tyr-57 in hSBP and Tyr-52 in rSBP). Removal of all three carbohydrate chains hSBP does not affect steroid binding activity (55,65). Human SBP binds DHT and E2 with Kds 0.4nM and 4nM, respectively, while rSBP binds DHT and E2 with Kds 0.8nM and 84nM, respectively (17). This indicates that the rabbit protein is essentially an androgen binding protein. This finding will be important for identifying E2 binding structural motifs (vide infra).

It became clear that continuing research on SBP would require training in molecular biology and cloning methods. I therefore took a sabbatical year leave of absence to learn these techniques. I was invited by Dr. Etienne Baulieu as a visiting professor at INSERM, Paris, in 1985. On my return to Seattle, I secured a collaboration with Dr. Fred Hagen of Zymogenetics and began the cloning and expression of hSBP-cDNA. We were not able to isolate a full-length clone (45) and decided to construct it from smaller fragments (54). It was expressed in mammalian cells (54, 56), insect cells (58), and in yeast (65). Site-directed mutagenesis revealed Tyr-57 (67), Met-107 (67), Lys-134 (56, 53), and Met-139 (56) as functional in steroid binding. Two of these residues (Tyr-57 and Lys-134) are at homologous positions in the rSBP sequence (Tyr-51 and Lys-128) but Met-107 is replaced with an isoleucine (Ileu-101 in rSBP). Since rSBP does not bind E2 significantly we initiated mutagenesis experiments to determine residues responsible for E2 binding in hSBP. The difference in the 3-D structures between DHT and E2 is small and resides mainly in the A-ring of the steroids, we therefore expected that replacement of not more than two residues would likely be sufficient to obtain a loss in E2 binding of hSBP. Aligning both sequences and starting from the amino terminus, 56 replacements were identified and combinatorial oligonucleotides coding for the changes were made for each selected segment. The results show that all products had wild type DHT Kd values and most of them had wild type E2 Kd values, except those having the R140K and I141L replacements (68). We conclude that R140 and I141 are located within a structural domain located of the steroid-binding site of hSBP where the A-ring of E2 is recognized.

While structural studies were advancing, research on the physiological role of SBP was initiated.  Availability of pure hSBP, nSBP, and monospecific hSBP antibodies (18), as well as collaboration with M. Novy and F. Stanczyk at the Oregon Primate Center allowed non-human primate studies. Clinical studies in the 1960s suggested that unbound steroids in human plasma diffused into tissues nonspecifically. With the discovery of SBP, it was proposed that it could function in controlling diffusion of sex steroids into tissues through the establishment of a steady-state equilibrium in plasma. There was support for this notion since the plasma metabolic clearance rate (MCR) of sex steroids in males and patients with low SBP levels was faster than normal and pregnant females which have higher levels of SBP. Infusion of pure nSBP and purified anti-SBP in the macaque supported this view by showing a direct effect of SBP on MCR-T (32, 51). The MCR-T decreased following the infusion of nSBP and increased following the infusion of pure hSBP antibodies. Furthermore, incubation of endometrial cancer cells in culture with pure hSBP inhibited the intracellular diffusion of E2 (34). We found that the effect of SBP on MCR-T is related to the distribution of T bound to albumin and SBP. The albumin plasma concentration does not vary but is much higher than SBP, but it binds T weakly and nonspecifically (Kd 5 x 10-5 M at 37oC) compared to SBP (Kd 4 x 10-9 M at 37oC). Calculations show that if % T bound to SBP and albumin is equal or higher than the % T bound to albumin alone, the influence on MCR-T is small. But if SBP is lowered such that T is mostly bound to albumin then there is a dramatic increase of MCR-T. There lies the role of SBP, it controls the level of “free” (unbound) steroid through a steady-state equilibrium with SBP and albumin. Albumin is absolutely required for this regulation. Fluctuation of plasma SBP levels therefore control the entry of sex steroids into tissues. Other roles for SBP have been suggested by us (24) and others, but there is insufficient data to support such proposals.

In 2000, I entered a collaboration with Wim Hol to crystallize the native and recombinant proteins for determining the 3-D structure of hSBP. We had expressed both the wild-type and fully-deglycosylated protein in yeast (65). In the meantime, a group in Germany published a crystal structure of the N-terminal domain of hSBP (residues 1 to 205) representing about 60% of the sequence (Grishkovskaya et al. EMBO J. 19:504-512, 2000). That partial sequence crystallized as a dimer and bound 2 moles DHT per mole dimer. The stoichiometry is different from that of the native protein (1 mole DHT/mole dimer) suggesting the possibility that the missing C-terminal domain (about 40% of the sequence) could function in negative cooperativity by masking the second potential binding site in the opposite subunit once the first site is occupied in the native dimer. The published model depicted all the residues we had predicted to be present in or near the steroid binding site (Tyr-57, Met-107, Met-139, R140, I-141), except Lys-134. That residue is not seen in the model because the published structure does not contain coordinates for residues 130 to 135. The most interesting finding came in determining the structural basis of E2 binding specificity. Recall that hSBP binds DHT and E2 whereas rSBP only binds the former. M107 is required for binding DHT and expression of both M107R and M107I mutants leaves E2 binding of hSBP intact (67). Eli Aldman, department of Biological Structure at UW, modeled the triple mutant M107I/R140K/I141L into the steroid binding site instead of the quadruple mutant M107I/I138V/R140K/I141L we had expressed, because the I138V had no effect on E2 binding. She found that there was no structural difference in the steroid binding site between wild-type SBP and the triple mutant when DHT is modeled into the site, consistent with the fact that a loss in DHT-binding affinity is not observed when these replacements are made (67). However, when E2 is modeled into the site containing the triple mutant a significant shift in structure occurred. Replacements at 140 and 141 with corresponding rSBP residues K and L, respectively, were sufficient to lower E2 binding affinity of hSBP without affecting the affinity of DHT. Interestingly, the side chains of R140 and I141 do not contact the steroid thus precluding a direct interaction as the explanation for differences in E2 affinity. Instead, it seems that the replacements affect losses of H-bonds in the network and changes in packing of E2 primarily with F56. That such small structural changes lead to significant changes in binding affinities makes sense in the evolution of the hSBP steroid-binding site. Because rSBP binds only T and DHT with high affinity, the ability of the human protein to bind all three steroids may have been a more recent evolutionary event. The data suggests that the mechanism for acquiring this new function involved the selection of subtle structural changes outside the immediate environment of the steroid binding site. Therefore, it seems that the steroid binding site of SBP was primarily designed to bind T and DHT and the ability to bind E2 was superimposed later onto the original structural design.

In 1999 and 2000 we repeatedly attempted to crystallize the native and full-length recombinant protein without success. I realized that continuing these efforts would not likely lead to the production of crystals, much less diffractable crystals. Having completed most of our research objectives I decided to close the lab in 2001. I kept my teaching responsibilities until 2006 at which time I fully retired. In closing, I want to acknowledge Hans Neurath and Walter Herrmann for giving me the opportunity of coming to Seattle, joining the faculty of the University of Washington, and contributing to its research mission. In addition, I want to thank Ken Walsh and Eli Adman for helping in understanding structure function relationships in SBP, and Fred Hagen for his involvement in the molecular biology.

Bibliography

  1. Petra P.H., Cohen W., Shaw E.N.: Isolation and characterization of the alkylated histidine from TLCK-inhibited trypsin. Biochem. Biophys. Res. Comm. 21, 612-618, 1965.
  2. Cohen W., and Petra P.H.: A kinetic analysis of the papain-catalyzed hydrolysis of a-N-benzoyl-l-citrulline methyl ester. Biochemistry 6, 1047-1053, 1967.
  3. Petra P.H., and Neurath H.: The heterogeneity of bovine carboxypeptidase A. I. The chromatographic purification of carboxypeptidase (Anson). Biochemistry 8, 2466-2475, 1969.
  4. Petra P.H., Bradshaw R.A., Walsh K.A., and Neurath H.: Identification of the allotype amino acid replacements of bovine carboxypeptidase A. Biochemistry 8, 2762-2768, 1969.
  5. Petra P.H., and Neurath H.: The heterogeneity of bovine carboxypeptidase A. II. The chromatographic purification of carboxypeptidase (Cox). Biochemistry 8, 5029-5036, 1969.
  6. Petra P.H.: The modification of carboxyl groups in bovine carboxypeptidase A. I. The inactivation of the enzyme by N-ethyl-5-phenylisoxazolium-3-sulfonate (Woodward’s Reagent K). Biochemistry 10, 3163-3170, 1971.
  7. Petra P.H., and Neurath H.: The modification of carboxyl groups in bovine carboxypeptidase A. II. Chemical identification of a functional glutamic acid residue and other reactive groups. Biochemistry 10, 3171-3177, 1971.
  8. Petra P.H., Hermodson M., Walsh K.A., and Neurath H.: Characterization of bovine carboxypeptidase A (Allan). Biochemistry 10, 4023-4025, 1971.
  9. Mickelson K.E., and Petra P.H.: A filter assay for the Sex Steroid-binding Protein (SBP) of human serum. FEBS Lett. 44, 34-38, 1974.
  10. Mickelson K.E., and Petra P.H.: Purification of the Sex Steroid-binding Protein (SBP) from human serum. Biochemistry 14, 957, 1975.
  11. Schiller H., and Petra P.H.: A filter assay for the corticosteroid-binding globulin (CBG) of human serum. J. Ster. Bioch. 7, 55-59, 1976.
  12. Gellert R.J., Lewis J., and Petra P.H.: Neonatal treatment with sex steroids. Relationship between the uterotropic response and the estrogen receptor in prepubertal rats. Endocrinology 100, 520-528, 1977.
  13. Petra P.H., and Schiller H.: Sex Steroid-binding Protein (SBP) in the plasma of the Macaca nemestrina. J. Ster. Bioch. 8, 655-661, 1977.
  14. Schiller H., Langley J.W., and Petra P.H.: Corticosteroid-binding protein in the plasma of the Macaca nemestrina. J. Ster. Bioch. 8, 647, 1977.
  15. Tabei T., Mickelson K.E., Neuhaus S., and Petra P.H.: Sex Steroid-binding protein (SBP) in dogs. J. Ster. Bioch. 9, 983-988, 1978.
  16. Mickelson K.E., Teller D.C., and Petra P.H. Characterization of the sex steroid binding protein of human pregnancy. Improvement in the purification procedure. Biochemistry 17, 1409-1415, 1978.
  17. Mickelson K.E., and Petra P.H.: Purification and physico-chemical characterization of the Sex Steroid-binding Protein of rabbit serum. Comparison with the human protein. J. Biol. Chem. 253, 5293-5298, 1978.
  18. Bordin S., Lewis J., and Petra P.H. : Monospecific antibody to the Sex Steroid-binding Protein (SBP) of human and rabbit serum: Cross-reactivity with other species. Bioch. Biophys. Res. Comm. 85, 391-401, 1978.
  19. Namkung P.C., Moe R.E., and Petra P.H.: Stability of estrogen receptors in frozen human breast tumor tissue. Cancer Res. 39, 1124-1125, 1979.
  20. Heinrichs W.L., Tabei T., Kuwubara Y., Burry K., Resko J., Petra P.H., Schiller H., and Namkung P.C.: Differentiation and regulation of peripheral androgen metabolism in rats and rhesus monkeys. Am. J. Obstet. Gynec. 135, 974-983, 1979.
  21. Raijfer J., Namkung P.C., and Petra P.H.: Ontogeny of androgen receptor in penis. Surgical Forum, Vol. XXX, Chapter SVII, 1979.
  22. Petra P.H., and Lewis J.: Modification in the purification of the sex steroid-binding protein of human serum by affinity chromatography. Anal. Bioch. 105, 165-169,1980.
  23. Burry K.A., Tabei T., Resko J., Petra P.H., and Heinrichs L.W.: Differentiation of sex steroid-binding protein in adult rhesus monkeys. Am. J. Obstet. Gyn. 136, 446-450, 1980.
  24. Bordin S., and Petra P.H.: Immunocytochemical localization of the plasma Sex Steroid-binding Protein (SBP) in tissues of the adult male monkey, Macaca nemestrina. Pro. Natl. Acad. Sci. (USA) 77, 5678-5682, 1980.
  25. Raijfer J., Namkung P.C., and Petra P.H.: Identification, partial characterization, and age-related changes of cytoplasmic androgen receptor in the rat penis. J. Steriod Bioch. 13, 1489-1492, 1980.
  26. Raijfer J., Namkung P.C., and Petra P.H.: The ontogeny of the cytoplasmic androgen receptor in the rat ventral prostate gland. Biol. Repro. 23, 518-521, 1980.
  27. Namkung P.C., and Petra P.H.: Measurement of progesterone receptors in human breast tumors. Comparison of various methods of analysis. J. Steroid Bioch. 14, 851-854, 1981.
  28. Bordin S., Torres R., and Petra P.H.: An enzyme immunoassay for the Sex steroid-binding Protein (SBP) of human plasma. J. Ster. Bioch. 17, 453-457, 1982.
  29. Ross J.B.A., Torres R., and Petra P.H.: Equilenin, a specific fluorescent probe for steroid-protein interactions in Sex Steroid-binding Protein (SBP). FEBS Letters 149, 240-244, 1982.
  30. Turner E., Ross J.B.A., Namkung P.C., and Petra P.H.: Purification and characterization of the sex steroid-binding protein from macaque serum. Comparison with the human protein. Biochemistry 23, 492-497, 1984.
  31. Stanczyk F.Z., Petra P.H., Senner J.W., and Novy M.J.: Effect of dexamethasone treatment on sex steroid-binding protein, corticosteroid-binding globulin, and steroid hormones in cycling rhesus macaques. Am. J. Obstet. Gyn. 151, 464-470, 1985.
  32. Petra P.H., Stanczyk F.Z., Namkung P.C., Fritz M.A., and Novy M.J.: Direct influence of the sex steroid-binding protein (SBP) of plasma on the metabolic clearance rate of testosterone. J. Steroid Bioch. 22, 739-746, 1985.
  33. David G.F., Koehler J.K., Brown J.A., Petra P.H., and Farr A.G.: Light and electron microscopic studies on the localization of the Sex Steroid-binding Protein (SBP) in rabbit spermatozoa. Biol. Reprod. 33, 503-514, 1985.
  34. De Ryck L., Ross J.B.A., Petra P.H., and Gurpide E.: Estradiol entry into endometrial cells in suspension. J. Ster. Bioch. 23, 145-152, 1985.
  35. Wortsman T., Frank S., Wehrenberg W.B., Petra P.H., and Murphy J.E.: Melanocyte-stimulating hormone immunoreactivity is a component of the neuroendocrine response to maximal stress (cardiac arrest). J. Clin. Endocr. Metab. 61, 355-360, 1985.
  36. Ross J.B.A., Contino P.B., Lulka M.F., and Petra P.H.: Observation and quantitation of metal binding sites in the Sex Steroid-binding Protein of human and rabbit sera using the luminescent probe, terbium. J. Prot. Chem. 4, 299-304, 1985.
  37. Petra P.H., Kumar S., Hayes R., Ericsson L.H., Titani K.: Molecular organization of the sex steroid-binding protein (SBP) of human plasma. J. Ster. Bioch. 24, 45-49, 1986.
  38. Orstan A., Lulka M.F., Eide B., Petra P.H., and Ross J.B.A.: Steroid-binding site of human and rabbit Sex Steroid-binding Protein of plasma: Fluorescence characterization with equilinin. Biochemistry 25, 2586-2692, 1986.
  39. Petra P.H., Namkung P.C., Senear D.F., McCrae D.A., Rousslang K.W., Teller D.C., and Ross J.B.A.: Molecular characterization of the sex steroid-binding protein (SBP) of plasma. Re-examination of rabbit SBP and comparison with the human, macaque, and baboon proteins. J. Ster. Bioch. 25, 191-200, 1986.
  40. Stanczyk F.Z., Hess D.L., Namkung P.C., Senner J.W., Petra P.H., and Novy M.J.: Alterations in Sex Steroid-binding Protein (SBP), corticosteroid binding globulin (CBG), and steroid hormone concentrations during pregnancy in rhesus macaques. Biol. Reprod. 35, 126-132, 1986.
  41. Walsh K.A., Titani K., Kumar S., Hayes R., and Petra P.H.: Amino acid sequence of the sex steroid-binding protein (SBP) of human blood plasma. Biochemistry 25, 7584-7590, 1986.
  42. Petra, P.H., Titani, K., Walsh, K.A., Joseph, D.R., Hall, S.H., and French, F.S. Comparison of the amino acid sequence of the sex steroid-binding protein of human plasma (SBP) with that of the androgen-binding protein (ABP) of rat testis, in: Binding Proteins of Steroid Hormones (Forest, M.G., and Pugeat, M. eds), , Colloque/INSERM, Vol. 149, pp. 137-142, 1986, John Libbey, London/Paris.
  43. Stanczyk F.Z., Namkung P.C., Fritz M.A., Novy M.J., and Petra P.H. The influence of sex steroid-binding protein on the metabolic clearance rate of testosterone. In: Binding Proteins of Steroid Hormones (Forest, M.G., and Pugeat, M. eds), Colloque/INSERM, Vol. 149, pp. 555-563, 1986, John Libbey, London/Paris.
  44. Petra P.H.: Measurement of the sex steroid-binding protein of human plasma by an enzyme-linked immunosorbent assay, ELISA. in: Binding Proteins of Steroid Hormones (Forest, M.G., and Pugeat, M. eds). Colloque/INSERM, Vol. 149, pp. 215-220, 1986, John Libbey, London/Paris.
  45. Que B.G., and Petra P.H.: Characterization of a cDNA coding for sex steroid-binding protein of human plasma. FEBS Letters 219: 405-409, 1987.
  46. Mercier-Bodard C., Radanyi C., Roux C., Groyer M.T., Robel P., Dadoune J.P., Petra P.H., Jolly D.J., and Baulieu E.E. (1987) Cellular distribution and hormonal regulation of hSBP in human hepatoma cells. J. Steroid Biochem. 27:297-307.
  47. Petra P.H., Que B., Namkung P.C., Ross J.B.A., Charbonneau H., Walsh K.A., Griffin P.R., Shabanowitz J., and Hunt D.F.: Affinity labeling, molecular cloning, and comparative amino acid sequence analysis of sex steroid-binding protein of plasma. A multidisciplinary approach for understanding steroid-protein interaction and its physiological role. Annals N.Y. Acad. Sci., Vol. 538, 10-24, 1988.
  48. Namkung P.C., Stanczyk F.Z., Cook M.J, Novy M.J., and Petra P.H. (1989) Half-life of Plasma Sex Steroid-binding Protein (SBP) in the Primate. J. Ster. Bioch. 32: 675-680 1989.
  49. Griffin R., Kumar S., Shabanowitz J., Charbonneau H., Namkung P.C., Walsh K.A., Hunt D.F., and Petra P.H. The amino acid sequence of the sex steroid-binding protein of rabbit serum. J. Biol. Chem. 264, 19066-19075, 1989.
  50. Plymate S.R, Namkung P.C., Matej L.A., and Petra P.H. Direct effect of plasma sex steroid-binding protein (SBP or SHBG) on the metabolic clearance rate of 17b-estradiol in the primate. J. Ster. Bioch. 36: 311-317, 1990.
  51. Casali E., Petra P.H., and Ross J.B.A. Fluorescence Investigation of the Sex steroid Binding Protein of Rabbit Serum: Steroid Binding and Subunit Dissociation. Biochemistry 29: 9334-9343, 1990.
  52. Namkung P.C., Kumar S., Walsh K.A., and Petra P.H. Identification of lysine-134 in the steroid-binding site of the sex steroid-binding protein of human plasma. J. Biol. Chem. 265: 18345-18350, 1990.
  53. Hagen F., Arguelles C., Sui L.M., Seidel P., Conroy S.C., & Petra P.H. Mammalian Expression of the Human Sex Steroid-Binding Protein of Plasma (SBP or SHBG) and Testis (ABP). Characterization of the Recombinant Protein. FEBS letters 299: 23-27 (1992).
  54. Petra PH, Griffin PR, and Moore K. Complete Enzymatic Deglycosylation of Native Sex Steroid-Binding Protein (SBP or SHBG) of Human and Rabbit Plasma. Effect on the Steroid-Binding Activity. Protein Science 1:902-909, 1992.
  55. Sui L.M., Cheung A.W.C., Namkung P.C., & Petra P.H. Localization of the Steroid-Binding Site of the Sex Steroid-Binding Protein (SBP or SHBG) of Human Plasma by Site-Directed Mutagenesis. FEBS Letters 310:115-118, 1992.
  56. Verlinde, C. L. M. J., Merritt, E. A., van den Akker, F., Kim, H., Fei, I., Delboni., L. F., Mande. S. C., Safarty. S., Petra, P. H., & Hol. W. G. J. Protein Crystallography and Infectious Diseases. Protein Science 3:1670-1686, 1994.
  57. Sui L.M., Wong C., & Petra P.H. Over-expression of Human Sex Steroid-Binding Protein (hSBP/hABP or hSHBG) in Insect Cells Infected with a Recombinant Baculovirus. Characterization of the Recombinant Protein and Comparison to the Plasma Protein. J. Steroid Biochem. Molec. Biol. 52:173-179, 1995.
  58. Kim, H., Feil, I., Verlinde, C. L. M. J., Petra, P. H., & Hol, W. G. J. Crystal Structure of Glycosomal Glyceraldehyde-3-Phosphate Dehydrogenase from Leishmania mexicana: Implications fro Structure-Based Drug Design and a New Position for the Inorganic Phosphate Binding Site. Biochemistry 34:14975-14986, 1995.
  59. Sui L.M., Hughes W., Hoppe A.J. & Petra P.H. Direct Evidence for the localization of the Steroid-binding site of the Plasma Sex Steroid-binding Protein (SBP or SHBG) at the Interface between the Subunits. Protein Science 5:2514-2520, 1996.
  60. Beck K., Gruber T., Ridgway C. C., Hughes W., Sui L.-M., and Petra, P.H. Secondary Structure and Shape of Plasma Sex Steroid-Binding Protein (SBP or SHBG). Comparison with Domain G of Laminin Results in a Structural Model of SBP. Eur. J. Biochem. 247:339-347 (1997)
  61. Bernstein B. E., Michels P.A.M., Kim H., Petra P.H., Hol W.G.J. The importance of dynamic light scattering in obtaining multiple crystals forms of T. brucei PGK. Protein Science 7:504-507 (1998)
  62. Mankoff D.A., Tewson T. J., Gralow J.R., Petra P.H., Peterson L.M., Woo I., Yaziji H., and Gown A.M. [18F]-16α-fluoroestradiol (FES) and positron emission tomography (PET) to measure estrogen receptor expression in breast cancer. Breast Ca. Res. Treat. 50:332 (1998).
  63. Sui L.M., Lennon J., Ma C., McCann I., Woo I., & Pétra P.H. Heterologous expression of wild type and deglycosylated human plasma sex steroid-binding protein (SBP or SHBG) in the yeast, Pichia pastoris. Characterization of recombinant proteins. J. Steroid. Biochem. Mol. Biol. 68:119-127. (1999)
  64. Tewson T. J., Mankoff D. A., Peterson L. M., Woo I., and Petra P. H. Interaction of 16a-[F-18]-Fluoroestradiol (FES) with Sex steroid Binding Protein (SBP) Nuclear Medicine & Biology 26:905-913 (1999)
  65. Pétra P. H., Woodcock K., Orr W. R., Nguyen D., and Sui L. M. The sex steroid binding protein (SBP or SHBG) of human plasma: Identification of Tyr-57 and Met-104 in the steroid binding site. J. Steroid Biochem. Mol. Biol. 75:139-145 (2000)
  66. Petra P.H., Adman E.T., Orr W.R., Woodcock K.T., Groff C., and Sui L-M. Arginine-140 and Isoleucine-141 determine the 17b-estradiol-binding specificity of the sex steroid-binding protein (SBP or SHBG) of human plasma. Protein Science 10: 1811-1821 (2001).
  67. Mankoff D.A., Peterson L.M., Tewson T. J., Stekhova S.A., Petra P.H., Gown A.M., Gralow J.R., Livingston R.B., Schubert E.K., and Krohn K.A., Non-invasive PET imaging of ER expression in breast cancer: sex steroid binding protein (SBP or SHBG) interaction and ER expression heterogeneity. Proceedings of the AACR 42:6 (2001).
  68. Linden H.M., Stekhova S.A., Link J.M., Gralow J.R., Livingston R.B., Ellis G.K., Petra P.H., Peterson L.M., Schubert E.K., Dunnwald L.K., Krohn K.A., and Mankoff D.A., Quantitative Fluoroestradiol Positron Emission Tomography Imaging Predicts Response to Endocrine Treatment in Breast Cancer. Journal Of Clinical Oncology 24: 2793-2799 (2006)

Reviews

  1. Neurath H., Bradshaw R.A., Ericsson L.H., Babin D.R., Petra P.H., Walsh K.A.: Current status of the chemical structure of bovine pancreatic carboxypeptidase A. Brookhaven Symposium in Biology 21, 1-23, 1968.
  2. Neurath H., Bradshaw R.A., Petra P.H., and Walsh K.A.: Bovine carboxypeptidase A. Activation, chemical structure, and molecular heterogeneity. Proc. Royal Soc. (London) B-257, 159-176, 1970.
  3. Petra P.H.: Bovine procarboxypeptidase and carboxypeptidase A. Methods in Enzymol. 19, 460-503, 1970.
  4. Petra P.H.: The serum sex steroid-binding protein. Purification, characterization, and immunological properties of the human and rabbit proteins. J. Ster. Bioch. 11, 245-252, 1979.
  5. Petra P.H., Stanczyk F.Z., Senear D.F., Namkung P.C., Novy M.J., Ross J.B.A., Turner E., and Brown J.A.: Current status of the molecular structure and function of the plasma sex steroid-binding protein (SBP). J. Ster. Bioch. 19, 699-706, 1983.
  6. Petra P.H., Namkung P.C., Titani K., and Walsh K.A. in: Binding Proteins of Steroid Hormones (Forest, M.G., and Pugeat, M., eds) Characterization of the plasma sex steroid-binding protein, Colloque/INSERM, Vol. 149, pp. 15-30, 1986, John Libbey, London/Paris.
  7. Petra, P.H. The Plasma Sex Steroid Binding Protein. A critical review of recent developments on the structure, molecular biology, and function. J. Steroid. Biochem. Mol. Biol. 40: 735-753 (1991).

Personal Data

I was born in Paris in 1937 and came to the United States in 1950 at the age of 13. My father, Yvon Petra, was a highly ranked tennis player in France before the war and reached a ranking of No. 4 in the world after winning Wimbledon in 1946 in singles, and the French Open in doubles in 1937 and 1946. In 2016 he was inducted posthumously into the International Tennis Hall of Fame in Newport, RI which I attended 1. He turned professional in 1947 and brought the family to the United Sates in 1950. I had learned tennis under his guidance in Belgium in 1947. I won the City of Chicago Tennis Championship in the Boys division (15 and under) in 1952, and reached the quarter finals of the United States Nationals Boys Tennis Championship in 1952 held in Kalamazoo, MI. In 1954, I reached the Finals of the Illinois State juniors Tennis Championship, and in 1959 won the New England Public Parks Open Tennis Championship in Hartford CT and reached the Finals of the Rhode Island Open State Tennis Championship in Providence. I went to Highschool in Palm Beach, Florida, and graduated in 1956. I was offered a tennis athletic scholarship at Tulane University in 1956 and graduated with a BS in chemistry i

n 1960. I attended Tulane University graduate school in 1960 and obtained an MS and PhD in biochemistry in 1966. Dolores and I became American citizens in 1962 and we were married that same year. in 1966, I was offered a postdoctoral position by Professor Hans Neurath, Chairman of the Biochemistry Department at the University of Washington. We moved to Seattle, and I continued playing tennis and won the Seattle City/Seafair Open Championship in 1968. I was offered a faculty position in the Department of Biochemistry and the Department of Ob/Gyn at the University of Washington in 1971. I stopped tennis competition but returned to it in 2001 at the time of my partial retirement.

1  https://www.newportri.com/story/sports/2016/07/21/after-hall-this-time/12778704007/

Selected Pictures

My Dad, my sister and I. Deauville, France, 1946.

My Dad, my sister and I. Deauville, France, 1946.

 

The College years: 1956-1960.
NCAA team champions, 1959. Tulane University, New Orleans.

 

The postdoctoral years: 1966-1970
Ralph and Penny Bradshaw, Dolores, and I. Bateaux Mouches, Paris 1969

 

The faculty years: 1970-2006
UW president McCormick visits my BIOC 406 class.

 

Organizing committee of the
First International Symposium
on steroid Binding Proteins.
Lyon France, 1986.

 

The sabbatical year: 1985
With Etienne Baulieu

 

The Retirement Years: 2006 – present
No more grant writing!