Crystal structure of active site mutant of antileukemic l -asparaginase reveals conserved zinc-binding site

Dominika Borek, Maciej Kozak, Jimin Pei, Mariusz Jaskolski

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

The periplasmic enzyme l-asparaginase type II from Escherichia coli (EcAII) converts l-asparagine to l-aspartate and ammonia. EcAII is an important drug in the treatment of childhood acute lymphoblastic leukemia, the most common malignancy in children. Leukemic blast cells lack the ability to synthesize l-asparagine and rely on other sources of l-asparagine for protein synthesis. EcAII injections deplete extracellular levels of l-asparagine, disrupting protein synthesis and inducing apoptosis in the malignant cells. The detailed mechanism of l-asparaginase catalytic action, the molecular mechanisms of its anticancer activity and the side effects associated with the treatment, including resistance to therapy, are not fully understood despite over 40 years of research. Here, we present X-ray structures of EcAII with an active site mutation, D90E, in three crystal forms. The region of the mutation is well ordered, allowing precise functional analysis of the consequences of the replacement of Asp90. In all three structures, the mutant protein exhibits an open conformation of the active site. In one of the structures, a zinc cation has been detected. The zinc cation is coordinated in a region of the protein that is implicated in the immunological response to EcAII treatment. A combined sequence-structure analysis of bacterial-type l-asparaginases reveals that the metal coordination may play a role in the response to asparaginase treatment. The observation of a zinc-binding site in antileukemic l-asparaginases provides new insight, with consequences for acute lymphoblastic leukemia therapy.

Original languageEnglish (US)
Pages (from-to)4097-4111
Number of pages15
JournalFEBS Journal
Volume281
Issue number18
DOIs
StatePublished - Sep 1 2014

Fingerprint

Asparaginase
Asparagine
Zinc
Catalytic Domain
Crystal structure
Binding Sites
Cations
Precursor Cell Lymphoblastic Leukemia-Lymphoma
Therapeutics
Functional analysis
Proteins
Bacterial Structures
Mutant Proteins
Mutation
Apoptosis Regulatory Proteins
Ammonia
Aspartic Acid
Escherichia coli
Conformations
Metals

Keywords

  • acute lymphoblastic leukemia
  • l -asparaginase
  • mutation
  • pancreatitis
  • zinc-binding site

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

Crystal structure of active site mutant of antileukemic l -asparaginase reveals conserved zinc-binding site. / Borek, Dominika; Kozak, Maciej; Pei, Jimin; Jaskolski, Mariusz.

In: FEBS Journal, Vol. 281, No. 18, 01.09.2014, p. 4097-4111.

Research output: Contribution to journalArticle

Borek, Dominika ; Kozak, Maciej ; Pei, Jimin ; Jaskolski, Mariusz. / Crystal structure of active site mutant of antileukemic l -asparaginase reveals conserved zinc-binding site. In: FEBS Journal. 2014 ; Vol. 281, No. 18. pp. 4097-4111.
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