5MQ5

A protease-resistant N24S Escherichia coli Asparaginase mutant with outstanding stability and enhanced anti-leukaemic activity

Summary for 5MQ5

• Entry DOI: 10.2210/pdb5mq5/pdb
• Descriptor: L-asparaginase 2, ASPARTIC ACID (3 entities in total)
• Functional Keywords: l-asparaginase, amidohydrolase, chemotherapeutic drug, hydrolase
• Biological source: Escherichia coli
• Total number of polymer chains: 4
• Total formula weight: 142247.05

Authors

Maggi, M.,Mittelman, S.D.,Parmentier, J.H.,Whitmire, J.M.,Merrell, D.S.,Scotti, C. (deposition date: 2016-12-20, release date: 2017-11-15, Last modification date: 2024-11-06)

Primary citation

Maggi, M.,Mittelman, S.D.,Parmentier, J.H.,Colombo, G.,Meli, M.,Whitmire, J.M.,Merrell, D.S.,Whitelegge, J.,Scotti, C.
A protease-resistant Escherichia coli asparaginase with outstanding stability and enhanced anti-leukaemic activity in vitro.
Sci Rep, 7:14479-14479, 2017

PubMed Abstract: L-Asparaginases (ASNases) have been used as first line drugs for paediatric Acute Lymphoblastic Leukaemia (ALL) treatment for more than 40 years. Both the Escherichia coli (EcAII) and Erwinia chrysanthemi (ErAII) type II ASNases currently used in the clinics are characterized by high in vivo instability, short half-life and the requirement of several administrations to obtain a pharmacologically active concentration. Moreover, they are sensitive to proteases (cathepsin B and asparagine endopeptidase) that are over-expressed by resistant leukaemia lymphoblasts, thereby impairing drug activity and pharmacokinetics. Herein, we present the biochemical, structural and in vitro antiproliferative characterization of a new EcAII variant, N24S. The mutant shows completely preserved asparaginase and glutaminase activities, long-term storage stability, improved thermal parameters, and outstanding resistance to proteases derived from leukaemia cells. Structural analysis demonstrates a modification in the hydrogen bond network related to residue 24, while Normal Mode-based geometric Simulation and Molecular Dynamics predict a general rigidification of the monomer as compared to wild-type. These improved features render N24S a potential alternative treatment to reduce the number of drug administrations in vivo and to successfully address one of the major current challenges of ALL treatment: spontaneous, protease-dependent and immunological inactivation of ASNase.

PubMed: 29101342
DOI: 10.1038/s41598-017-15075-4

Experimental method

X-RAY DIFFRACTION (1.6 Å)