Crystal structure of carboxypeptidase A complexed with an inactivator in two crystal forms

Protein Eng. 1998 Oct;11(10):841-5. doi: 10.1093/protein/11.10.841.

Abstract

Two different crystal forms of carboxypeptidase A (CPA) complexed with an inactivator were obtained by the method of hanging drop vapor diffusion. The inactivator, 2-benzyl-3-iodo-propanoic acid (BIPA), binds covalently to an active site residue Glu270 of CPA. The complexes were crystallized in the space group P2(1) (CPA-I) and P2(1)2(1)2(1) (CPA-II), respectively. The structures of both crystal forms were determined by molecular replacement using the native CPA crystal structure as the search model. The final crystallographic residuals are 0.163 for CPA-I and 0.152 for CPA-II. Except for the modification of Glu270, the inactivator exhibits normal binding mode compared with other ligand complexes of CPA. In the final electron density difference maps (2Fo-Fc, Fo-Fc), the density of the iodo ion could not be found in both crystal forms while the conserved water molecule remains coordinated to Zn2+ as in the native CPA. Comparisons of the complexes of CPA-BIPA with the native CPA and the CPA-D-Phe complex are presented. The mechanism of the inactivation of CPA and its implication for catalytic mechanism were discussed.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Benzene Derivatives
  • Binding Sites
  • Carboxypeptidases / antagonists & inhibitors
  • Carboxypeptidases / chemistry*
  • Carboxypeptidases / metabolism
  • Carboxypeptidases A
  • Cattle
  • Crystallization
  • Crystallography, X-Ray
  • Electrons
  • Glutamic Acid / metabolism
  • Hydrogen-Ion Concentration
  • Ligands
  • Models, Chemical
  • Pancreas
  • Propionates / metabolism*
  • Protein Conformation
  • Temperature
  • Water / metabolism
  • Zinc / metabolism

Substances

  • 2-benzyl-3-iodo-propanoic acid
  • Benzene Derivatives
  • Ligands
  • Propionates
  • Water
  • Glutamic Acid
  • Carboxypeptidases
  • Carboxypeptidases A
  • Zinc