PDBsum entry 2ewb

Hydrolase

PDB id: 2ewb

Contents

Protein chain

486 a.a. *

Ligands

CO3

ZED

MPD ×4

Metals

_ZN ×2

_NA

Waters ×477

* Residue conservation analysis

PDB id:

2ewb

Name:

Hydrolase

Title:

The crystal structure of bovine lens leucine aminopeptidase in complex with zofenoprilat

Structure:

Cytosol aminopeptidase. Chain: a. Synonym: leucine aminopeptidase, lap, leucyl aminopeptidase, proline aminopeptidase. Ec: 3.4.11.1

Source:

Bos taurus. Cattle. Organism_taxid: 9913. Other_details: lens

Biol. unit:

Hexamer (from PDB file)

Resolution:

1.85Å R-factor: 0.156 R-free: 0.177

Authors:

V.Alterio,C.Pedone,G.De Simone

Key ref:

M.Cappiello et al. (2006). Metal ion substitution in the catalytic site greatly affects the binding of sulfhydryl-containing compounds to leucyl aminopeptidase. Biochemistry, 45, 3226-3234. PubMed id: 16519517 DOI: 10.1021/bi052069v

Date:

02-Nov-05 Release date: 04-Apr-06

Protein chain

P00727  (AMPL_BOVIN) -  Cytosol aminopeptidase from Bos taurus

Seq: 519 a.a.

Struc: 486 a.a.

Enzyme reactions

• Enzyme class 1: E.C.3.4.11.1 - leucyl aminopeptidase.
• Reaction: Release of an N-terminal amino acid, Xaa-|-Xbb-, in which Xaa is preferably Leu, but may be other amino acids including Pro although not Arg or Lys, and Xbb may be Pro.
• Cofactor: Zn(2+)
• Enzyme class 2: E.C.3.4.11.5 - prolyl aminopeptidase.
• Reaction: Release of a N-terminal proline from a peptide.
• Cofactor: Mn(2+)
• Enzyme class 3: E.C.3.4.13.23 - cysteinylglycine-S-conjugate dipeptidase.
• Reaction: an S-substituted L-cysteinylglycine + H2O = an S-substituted L-cysteine + glycine

S-substituted L-cysteinylglycine + H2O = S-substituted L-cysteine + glycine (Bound ligand (Het Group name = CO3) matches with 50.00% similarity)

Note, where more than one E.C. class is given (as above), each may correspond to a different protein domain or, in the case of polyprotein precursors, to a different mature protein.

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

Added reference

DOI no: 10.1021/bi052069v

Biochemistry 45:3226-3234 (2006)

PubMed id: 16519517

Metal ion substitution in the catalytic site greatly affects the binding of sulfhydryl-containing compounds to leucyl aminopeptidase.

M.Cappiello, V.Alterio, P.Amodeo, A.Del Corso, A.Scaloni, C.Pedone, R.Moschini, G.M.De Donatis, G.De Simone, U.Mura.

ABSTRACT

Bovine lens leucyl aminopeptidase (blLAP), a homohexameric metallopeptidase preferring bulky and hydrophobic amino acids at the N-terminus of (di)peptides, contains two Zn(2+) ions per subunit that are essential for catalytic activity. They may be replaced by other divalent cations with different exchange kinetics. The protein readily exchangeable site (site 1) can be occupied by Zn(2+), Mn(2+), Mg(2+), or Co(2+), while the tight binding site (site 2) can be occupied by Zn(2+) or Co(2+). We recently reported that introduction of Mn(2+) into site 1 generates a novel activity of blLAP toward CysGly [Cappiello, M., et al. (2004) Biochem. J. 378, 35-44], which in contrast is not hydrolyzed by the (Zn/Zn) enzyme. This finding, while disclosing a potential specific role for blLAP in glutathione metabolism, raised a question about the features required for molecules to be a substrate for the enzyme. To clarify the interaction of the enzyme with sulfhydryl-containing derivatives, (Zn/Zn)- and (Mn/Zn)blLAP forms were prepared and functional-structural studies were undertaken. Thus, a kinetic analysis of various compounds with both enzyme forms was performed; the crystal structure of (Zn/Zn)blLAP in complex with the peptidomimetic derivative Zofenoprilat was determined, and a modeling study on the CysGly-(Zn/Zn)blLAP complex was carried out. This combined approach provided insight into the interaction of blLAP with sulfhydryl-containing derivatives, showing that the metal exchange in site 1 modulates binding to these molecules that may result in enzyme substrates or inhibitors, depending on the nature of the metal.