PDBsum entry 3be7

Hydrolase

PDB id: 3be7

Contents

Protein chains

(+ 2 more) 398 a.a. *

Ligands

ARG ×8

GOL ×16

Metals

_MG ×2

Waters ×572

* Residue conservation analysis

PDB id:

3be7

Name:

Hydrolase

Title:

Crystal structure of zn-dependent arginine carboxypeptidase

Structure:

Zn-dependent arginine carboxypeptidase. Chain: a, b, c, d, e, f, g, h. Engineered: yes

Source:

Unidentified. Organism_taxid: 32644. Gene: ibea_ctg=2118574. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008. Other_details: environmental sample from sargasso sea

Resolution:

2.30Å R-factor: 0.228 R-free: 0.280

Authors:

Y.Patskovsky,U.A.Ramagopal,R.Toro,A.J.Meyer,J.Freeman,M.Iizuka, K.Bain,L.Rodgers,F.Raushel,J.M.Sauder,S.K.Burley,S.C.Almo,New York Sgx Research Center For Structural Genomics (Nysgxrc)

Key ref:

D.F.Xiang et al. (2009). Functional identification of incorrectly annotated prolidases from the amidohydrolase superfamily of enzymes. Biochemistry, 48, 3730-3742. PubMed id: 19281183

Date:

16-Nov-07 Release date: 27-Nov-07

Protein chains

No UniProt id for this chain

Struc: 398 a.a.

Biochemistry 48:3730-3742 (2009)

PubMed id: 19281183

Functional identification of incorrectly annotated prolidases from the amidohydrolase superfamily of enzymes.

D.F.Xiang, Y.Patskovsky, C.Xu, A.J.Meyer, J.M.Sauder, S.K.Burley, S.C.Almo, F.M.Raushel.

ABSTRACT

The substrate profiles for two proteins from Caulobacter crescentus CB15 (Cc2672 and Cc3125) and one protein (Sgx9359b) derived from a DNA sequence ( gi|44368820 ) isolated from the Sargasso Sea were determined using combinatorial libraries of dipeptides and N-acyl derivatives of amino acids. These proteins are members of the amidohydrolase superfamily and are currently misannotated in NCBI as catalyzing the hydrolysis of l-Xaa-l-Pro dipeptides. Cc2672 was shown to catalyze the hydrolysis of l-Xaa-l-Arg/Lys dipeptides and the N-acetyl and N-formyl derivatives of lysine and arginine. This enzyme will also hydrolyze longer peptides that terminate in either lysine or arginine. The N-methyl phosphonate derivative of l-lysine was a potent competitive inhibitor of Cc2672 with a K(i) value of 120 nM. Cc3125 was shown to catalyze the hydrolysis of l-Xaa-l-Arg/Lys dipeptides but will not hydrolyze tripeptides or the N-formyl and N-acetyl derivatives of lysine or arginine. The substrate profile for Sgx9359b is similar to that of Cc2672 except that compounds with a C-terminal lysine are not recognized as substrates. The X-ray structure of Sgx9359b was determined to a resolution of 2.3 A. The protein folds as a (beta/alpha)(8)-barrel and self-associates to form a homooctamer. The active site is composed of a binuclear metal center similar to that found in phosphotriesterase and dihydroorotase. In one crystal form, arginine was bound adventitiously to the eight active sites within the octamer. The orientation of the arginine in the active site identified the structural determinants for recognition of the alpha-carboxylate and the positively charged side chains of arginine-containing substrates. This information was used to identify 18 other bacterial sequences that possess identical or similar substrate profiles.