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PMID: 3863130 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Binding of a possible transition state analogue to the active site of carboxypeptidase A.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 82 ·No. 20 ·1985-10-00 ·Pages 6840-4

Christianson DW, Lipscomb WN

Abstract

The mode of binding of the competitive inhibitor 2-benzyl-3-formylpropanoic acid to the active site of carboxypeptidase A has been studied by x-ray diffraction methods to a resolution of 1.7 A. The actual species bound to the enzyme was determined to be the gem-diol resulting from covalent hydration at the aldehyde carbonyl. Details relating to the process of association of inhibitor with enzyme are unknown at this time: the free aldehyde could initially bind to the enzyme and subsequently undergo catalytic hydration; or, the hydrate itself could be the species initially binding to the enzyme, because it does exist to a high degree (25%) in aqueous solution. Nevertheless, the structure of the complex reported is reminiscent of a possible tetrahedral intermediate that would be encountered in a general base hydrolytic mechanism. Of course, other mechanistic proposals, such as the anhydride pathway, cannot be ruled out simply on the basis of the structure of this enzyme-inhibitor complex.

MeSH Terms
Binding Sites Binding, Competitive Carboxypeptidases/metabolism Carboxypeptidases A Kinetics Models, Molecular Phenylpropionates/pharmacology Protein Binding Protein Conformation X-Ray Diffraction
Chemicals
Phenylpropionates 2-benzyl-3-formylpropanoic acid Carboxypeptidases Carboxypeptidases A
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Christianson D W
Lipscomb W N
References (39)
39 references, click to expand
  1. Crystallographic studies on apocarboxypeptidase A and the complex with glycyl-L-tyrosine.
    Proc Natl Acad Sci U S A. 1983 Dec;80(23):7151-4 PMID: 6580631
  2. Transition state analogues for enzyme catalysis.
    Nature. 1969 Aug 16;223(5207):704-5 PMID: 4979456
  3. Structure of the potato inhibitor complex of carboxypeptidase A at 2.5-A resolution.
    Proc Natl Acad Sci U S A. 1980 Aug;77(8):4633-7 PMID: 6933511
  4. Aldehydes as inhibitors of papain.
    J Biol Chem. 1972 Dec 25;247(24):8195-7 PMID: 4640942
  5. Crystallographic and kinetic investigations of the covalent complex formed by a specific tetrapeptide aldehyde and the serine protease from Streptomyces griseus.
    Proc Natl Acad Sci U S A. 1979 Jan;76(1):96-100 PMID: 106392
  6. The metallobiochemistry of zinc enzymes.
    Adv Enzymol Relat Areas Mol Biol. 1984;56:283-430 PMID: 6364704
  7. CRYSTALLINE CARBOXYPOLYPEPTIDASE.
    Science. 1935 May 10;81(2106):467-8 PMID: 17818766
  8. Similarities between the conformation of arsanilazotyrosine 248 of carboxypeptidase A in the crystalline state and in solution.
    Proc Natl Acad Sci U S A. 1972 Oct;69(10):2850-4 PMID: 4507609
  9. Mechanism of action of carboxypeptidase A in ester hydrolysis.
    Proc Natl Acad Sci U S A. 1976 Nov;73(11):3882-6 PMID: 1069272
  10. Use of secondary isotope effects and varying pH to investigate the mode of binding of inhibitory amino aldehydes by leucine aminopeptidase.
    Biochemistry. 1985 Jan 15;24(2):330-3 PMID: 3978076
  11. Enzymatic catalysis and transition-state theory.
    Science. 1973 Apr 15;180(4082):149-54 PMID: 4632837
  12. Binding of ligands to the active site of carboxypeptidase A.
    Proc Natl Acad Sci U S A. 1981 Sep;78(9):5455-9 PMID: 6946483
  13. Transition-state analogs of an aliphatic amidase.
    FEBS Lett. 1973 Sep 1;35(1):109-11 PMID: 4201665
  14. Binding of N-carboxymethyl dipeptide inhibitors to thermolysin determined by X-ray crystallography: a novel class of transition-state analogues for zinc peptidases.
    Biochemistry. 1984 Nov 20;23(24):5724-9 PMID: 6395881
  15. Rat preprocarboxypeptidase A: cDNA sequence and preliminary characterization of the gene.
    Proc Natl Acad Sci U S A. 1982 Jan;79(1):31-5 PMID: 6275388
  16. Use of peptide aldehydes to generate transition-state analogs of elastase.
    Biochemistry. 1973 Jan 2;12(1):47-51 PMID: 4734224
  17. Refined crystal structure of carboxypeptidase A at 1.54 A resolution.
    J Mol Biol. 1983 Aug 5;168(2):367-87 PMID: 6887246
  18. The amino acid sequence of bovine carboxypeptidase A.
    Proc Natl Acad Sci U S A. 1969 Aug;63(4):1389-94 PMID: 5260942
  19. Mutagenesis at a specific position in a DNA sequence.
    J Biol Chem. 1978 Sep 25;253(18):6551-60 PMID: 681366
  20. Enzymatic catalysis and the transition state theory of reaction rates: transition state analogs.
    Cold Spring Harb Symp Quant Biol. 1972;36:45-51 PMID: 4508159
  21. Aspartic-beta-semialdehyde: a potent inhibitor of Escherichia coli L-asparaginase.
    J Biol Chem. 1974 Oct 10;249(19):6351-3 PMID: 4609075
  22. Refined crystal structure of the potato inhibitor complex of carboxypeptidase A at 2.5 A resolution.
    J Mol Biol. 1982 Sep 25;160(3):475-98 PMID: 7154070
  23. Inhibition of carboxypeptidase A by aldehyde and ketone substrate analogues.
    Biochemistry. 1984 Apr 24;23(9):2083-7 PMID: 6547054
  24. Peptide aldehydes inhibiting chymotrypsin.
    Biochem Biophys Res Commun. 1972 Oct 17;49(2):343-9 PMID: 4640362
  25. The structure of carboxypeptidase A. VII. The 2.0-angstrom resolution studies of the enzyme and of its complex with glycyltyrosine, and mechanistic deductions.
    Brookhaven Symp Biol. 1968 Jun;21(1):24-90 PMID: 5719196
  26. Design of a diffractometer and flow cell system for X-ray analysis of crystalline proteins with applications to the crystal chemistry of ribonuclease-S.
    J Mol Biol. 1967 Aug 14;27(3):563-78 PMID: 6049685
  27. The sequence around the active-center tyrosyl residue of bovine pancreatic carboxypeptidase A.
    Proc Natl Acad Sci U S A. 1967 Jul;58(1):280-5 PMID: 5231609
  28. Structure of an actively exchanging complex between carboxypeptidase A and a substrate analogue.
    Proc Natl Acad Sci U S A. 1980 Jun;77(6):3288-91 PMID: 6932021
  29. Thiohemiacetal formation by inhibitory aldehydes at the active site of papain.
    Biochemistry. 1977 Nov 1;16(22):4890-5 PMID: 911798
  30. INTERMOLECULAR CROSS LINKING OF A PROTEIN IN THE CRYSTALLINE STATE: CARBOXYPEPTIDASE-A.
    Proc Natl Acad Sci U S A. 1964 Sep;52:833-9 PMID: 14212562
  31. A peptidase-inactive derivative of carboxypeptidase A modified specifically at tyrosine 248. Cobalt(III) (ethylenediamine-N,N'-diacetato) (arsanilazotyrosinato 248 carboxypeptidase A).
    J Biol Chem. 1979 Dec 10;254(23):11868-74 PMID: 574142
  32. Structure and catalysis of enzymes.
    Annu Rev Biochem. 1983;52:17-34 PMID: 6225375
  33. alpha-aminoaldehydes: transition state analogue inhibitors of leucine aminopeptidase.
    Biochemistry. 1982 Aug 17;21(17):4177-80 PMID: 7126535
  34. Catalytic role of the metal ion of carboxypeptidase A in ester hydrolysis.
    J Biol Chem. 1979 Jan 25;254(2):356-66 PMID: 33168
  35. Carboxypeptidase A: a protein and an enzyme.
    Adv Protein Chem. 1971;25:1-78 PMID: 4946703
  36. Carboxypeptidase A mechanisms.
    Proc Natl Acad Sci U S A. 1980 Jul;77(7):3875-8 PMID: 6933442
  37. PROCEDURES FOR THE ISOLATION OF CRYSTALLINE BOVINE PANCREATIC CARBOXYPEPTIDASE A. II. ISOLATION OF CARBOXYPEPTIDASE A-ALPHA FROM PROCARBOXYPEPTIDASE A.
    Biochemistry. 1964 Jan;3:44-7 PMID: 14114502
  38. Zinc environment and cis peptide bonds in carboxypeptidase A at 1.75-A resolution.
    Proc Natl Acad Sci U S A. 1981 Jun;78(6):3408-12 PMID: 6943549
  39. Transition state analog inhibitors and enzyme catalysis.
    Annu Rev Biophys Bioeng. 1976;5:271-306 PMID: 7991
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.82.20.6840
Published
1985-10-00
Pages
6840-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC390783
Subset
IM
Grants
NIGMS NIH HHS · GM 06920 · United States
External Links
PubMed source DOI: 10.1073/pnas.82.20.6840 PMC Full Text View journal details
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