Substrate specificity of prostate-specific antigen (PSA)

Gary S. Coombs, Robert C. Bergstrom, Jean Luc Pellequer, Scott I. Baker, Marc Navre, Matthew M. Smith, John A. Tainer, Edwin L. Madison, David R. Corey

Research output: Contribution to journalArticle

105 Citations (Scopus)

Abstract

Background: The serine protease prostate-specific antigen (PSA) is a useful clinical marker for prostatic malignancy. PSA is a member of the kallikrein subgroup of the (chymo)trypsin serine protease family, but different from the prototypical member of this subgroup, tissue kallikrein, in possessing a specificity more similar to that of chymotrypsin than trypsin. We report the use of two strategies, substrate phage display and iterative optimization of natural cleavage sites, to identify labile sequences for PSA cleavage. Results: Iterative optimization and substrate phage display converged on the amino-acid sequence SS(Y/F)Y↓(G/S) as preferred subsite occupancy for PSA. These sequences were cleaved by PSA with catalytic efficiencies as high as 2200-3100 M-1 s-1, compared with values of 2-46 M-1 s-1 for peptides containing likely physiological target sequences of PSA from the protein semenogelin. Substrate residues that bind to secondary (non-S1) subsites have a critical role in defining labile substrates and can even cause otherwise disfavored amino acids to bind in the primary specificity (S1) pocket. Conclusions: The importance of secondary subsites in defining both the specificity and efficiency of cleavage suggests that substrate recognition by PSA is mediated by an extended binding site. Elucidation of preferred subsite occupancy allowed refinement of the structural model of PSA and should facilitate the development of more sensitive activity-based assays and the design of potent inhibitors.

Original languageEnglish (US)
Pages (from-to)475-488
Number of pages14
JournalChemistry and Biology
Volume5
Issue number9
StatePublished - Sep 1998

Fingerprint

Prostate-Specific Antigen
Substrate Specificity
Substrates
Bacteriophages
Serine Proteases
Display devices
Tissue Kallikreins
Amino Acids
Kallikreins
Structural Models
Trypsin
Amino Acid Sequence
Assays
Biomarkers
Binding Sites
Peptides
Neoplasms

Keywords

  • Prostate-specific antigen
  • Protease specificity
  • Subsite occupancy
  • Substrate phage display

ASJC Scopus subject areas

  • Organic Chemistry

Cite this

Coombs, G. S., Bergstrom, R. C., Pellequer, J. L., Baker, S. I., Navre, M., Smith, M. M., ... Corey, D. R. (1998). Substrate specificity of prostate-specific antigen (PSA). Chemistry and Biology, 5(9), 475-488.

Substrate specificity of prostate-specific antigen (PSA). / Coombs, Gary S.; Bergstrom, Robert C.; Pellequer, Jean Luc; Baker, Scott I.; Navre, Marc; Smith, Matthew M.; Tainer, John A.; Madison, Edwin L.; Corey, David R.

In: Chemistry and Biology, Vol. 5, No. 9, 09.1998, p. 475-488.

Research output: Contribution to journalArticle

Coombs, GS, Bergstrom, RC, Pellequer, JL, Baker, SI, Navre, M, Smith, MM, Tainer, JA, Madison, EL & Corey, DR 1998, 'Substrate specificity of prostate-specific antigen (PSA)', Chemistry and Biology, vol. 5, no. 9, pp. 475-488.
Coombs GS, Bergstrom RC, Pellequer JL, Baker SI, Navre M, Smith MM et al. Substrate specificity of prostate-specific antigen (PSA). Chemistry and Biology. 1998 Sep;5(9):475-488.
Coombs, Gary S. ; Bergstrom, Robert C. ; Pellequer, Jean Luc ; Baker, Scott I. ; Navre, Marc ; Smith, Matthew M. ; Tainer, John A. ; Madison, Edwin L. ; Corey, David R. / Substrate specificity of prostate-specific antigen (PSA). In: Chemistry and Biology. 1998 ; Vol. 5, No. 9. pp. 475-488.
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