2JK0

Structural and functional insights into Erwinia carotovora L- asparaginase

Summary for 2JK0

• Entry DOI: 10.2210/pdb2jk0/pdb
• Related: 2VM7
• Descriptor: L-ASPARAGINASE, ASPARTIC ACID (3 entities in total)
• Functional Keywords: erwinia, hydrolase, enzyme therapy, protein stability, leukemia treatment
• Biological source: PECTOBACTERIUM CAROTOVORUM
• Total number of polymer chains: 8
• Total formula weight: 275689.42

Authors

Papageorgiou, A.C.,Posypanova, G.A.,Andersson, C.S.,Sokolov, N.N.,Krasotkina, J. (deposition date: 2008-05-23, release date: 2008-08-05, Last modification date: 2023-12-13)

Primary citation

Papageorgiou, A.C.,Posypanova, G.A.,Andersson, C.S.,Sokolov, N.N.,Krasotkina, J.
Structural and Functional Insights Into Erwinia Carotovora L-Asparaginase.
FEBS J., 275:4306-, 2008

PubMed 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.

PubMed: 18647344
DOI: 10.1111/J.1742-4658.2008.06574.X

Experimental method

X-RAY DIFFRACTION (2.5 Å)