PDBsum entry: 2jk0

Hydrolase

PDB id: 2jk0

Contents

Protein chains

(+ 0 more) 325 a.a. *

309 a.a. *

Ligands

ASP ×8

Waters ×708

* Residue conservation analysis

PDB id:

2jk0

Name:

Hydrolase

Title:

Structural and functional insights into erwinia carotovora l-asparaginase

Structure:

L-asparaginase. Chain: a, b, c, d, e, f, g, h. Engineered: yes

Source:

Pectobacterium carotovorum. Organism_taxid: 554. Expressed in: escherichia coli. Expression_system_taxid: 469008.

Resolution:

2.50Å R-factor: 0.192 R-free: 0.266

Authors:

A.C.Papageorgiou,G.A.Posypanova,C.S.Andersson,N.N.Sokolov, J.Krasotkina

Key ref:

A.C.Papageorgiou et al. (2008). Structural and functional insights into Erwinia carotovora l-asparaginase. Febs J, 275, 4306-4316. PubMed id: 18647344 DOI: 10.1111/j.1742-4658.2008.06574.x

Date:

23-May-08 Release date: 05-Aug-08

Protein chains

Q6Q4F4  (Q6Q4F4_ERWCT) -  L-asparaginase (Precursor)

Seq: 346 a.a.

Struc: 325 a.a.

Protein chains

Q6Q4F4  (Q6Q4F4_ERWCT) -  L-asparaginase (Precursor)

Seq: 346 a.a.

Struc: 309 a.a.

Enzyme reactions

• Enzyme class: Chains A, B, C, D, E, F, G, H: E.C.3.5.1.1 - Asparaginase.
• Reaction: L-asparagine + H₂O = L-aspartate + NH₃

L-asparagine + H(2)O = L-aspartate (Bound ligand (Het Group name = ASP) corresponds exactly) + NH(3)

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

 Gene Ontology (GO) functional annotation 

• Biological process: cellular amino acid metabolic process (2 terms)
• Biochemical function: hydrolase activity (2 terms)

Added reference

DOI no: 10.1111/j.1742-4658.2008.06574.x

Febs J 275:4306-4316 (2008)

PubMed id: 18647344

Structural and functional insights into Erwinia carotovora l-asparaginase.

A.C.Papageorgiou, G.A.Posypanova, C.S.Andersson, N.N.Sokolov, J.Krasotkina.

ABSTRACT

Bacterial l-asparaginases are enzymes that catalyze the hydrolysis of l-asparagine to aspartic acid. For the past 30 years, these enzymes have been used as therapeutic agents in the treatment of acute childhood lymphoblastic leukemia. Their intrinsic low-rate glutaminase activity, however, causes serious side-effects, including neurotoxicity, hepatitis, coagulopathy, and other dysfunctions. Erwinia carotovora asparaginase shows decreased glutaminase activity, so it is believed to have fewer side-effects in leukemia therapy. To gain detailed insights into the properties of E. carotovora asparaginase, combined crystallographic, thermal stability and cytotoxic experiments were performed. The crystal structure of E. carotovoral-asparaginase in the presence of l-Asp was determined at 2.5 A resolution and refined to an R(cryst) of 19.2 (R(free) = 26.6%) with good stereochemistry. Cytotoxicity measurements revealed that E. carotovora asparaginase is 30 times less toxic than the Escherichia coli enzyme against human leukemia cell lines. Moreover, denaturing experiments showed that E. carotovora asparaginase has decreased thermodynamic stability as compared to the E. coli enzyme and is rapidly inactivated in the presence of urea. On the basis of these results, we propose that E. carotovora asparaginase has limited potential as an antileukemic drug, despite its promising low glutaminase activity. Our analysis may be applicable to the therapeutic evaluation of other asparaginases as well.