当前位置： SCI文献检索 > BIOCHEMISTRY期刊下所有文献 > Structural basis for the activity and substrate specificity of Erwinia chrysanthemi L-asparaginase.

Structural basis for the activity and substrate specificity of Erwinia chrysanthemi L-asparaginase.

Abstract：

:Bacterial L-asparaginases, enzymes that catalyze the hydrolysis of L-asparagine to aspartic acid, have been used for over 30 years as therapeutic agents in the treatment of acute childhood lymphoblastic leukemia. Other substrates of asparaginases include L-glutamine, D-asparagine, and succinic acid monoamide. In this report, we present high-resolution crystal structures of the complexes of Erwinia chrysanthemi L-asparaginase (ErA) with the products of such reactions that also can serve as substrates, namely L-glutamic acid (L-Glu), D-aspartic acid (D-Asp), and succinic acid (Suc). Comparison of the four independent active sites within each complex indicates unique and specific binding of the ligand molecules; the mode of binding is also similar between complexes. The lack of the alpha-NH3(+) group in Suc, compared to L-Asp, does not affect the binding mode. The side chain of L-Glu, larger than that of L-Asp, causes several structural distortions in the ErA active side. The active site flexible loop (residues 15-33) does not exhibit stable conformation, resulting in suboptimal orientation of the nucleophile, Thr15. Additionally, the delta-COO(-) plane of L-Glu is approximately perpendicular to the plane of gamma-COO(-) in L-Asp bound to the asparaginase active site. Binding of D-Asp to the ErA active site is very distinctive compared to the other ligands, suggesting that the low activity of ErA against D-Asp could be mainly attributed to the low k(cat) value. A comparison of the amino acid sequence and the crystal structure of ErA with those of other bacterial L-asparaginases shows that the presence of two active-site residues, Glu63(ErA) and Ser254(ErA), may correlate with significant glutaminase activity, while their substitution by Gln and Asn, respectively, may lead to minimal L-glutaminase activity.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Aghaiypour K,Wlodawer A,Lubkowski J

doi

10.1021/bi0029595

subject

Has Abstract

pub_date

2001-05-15 00:00:00

pages

5655-64

issue

19

eissn

0006-2960

issn

1520-4995

pii

bi0029595

journal_volume

40

pub_type

杂志文章

相关文献

BIOCHEMISTRY文献大全