PT  - JOURNAL ARTICLE
AU  - Talita Stelling de Araújo
AU  - Sandra M. N. Scapin
AU  - William de Andrade
AU  - Maira Fasciotti
AU  - Mariana T. Q. de Magalhães
AU  - Marcius S. Almeida
AU  - Luís Maurício T. R. Lima
TI  - Biophysical characterization of two commercially available preparations of the drug containing <em>Escherichia coli</em> L-Asparaginase 2
AID  - 10.1101/2020.11.11.379065
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.11.11.379065
4099  - http://biorxiv.org/content/early/2020/11/12/2020.11.11.379065.short
4100  - http://biorxiv.org/content/early/2020/11/12/2020.11.11.379065.full
AB  - The hydrolysis of asparagine and glutamine by L-asparaginase has been used to treat acute lymphoblastic leukemia for over four decades. Each L-asparaginase monomer has a long loop that closes over the active site upon substrate binding, acting as a lid. Here we present a comparative study two commercially available preparations of the drug containing Escherichia coli L-Asparaginase 2, performed by a comprehensive array of biophysical and biochemical approaches. We report the oligomeric landscape and conformational and dynamic plasticity of E. coli type 2 L-asparaginase (EcA2) present in two different formulations, and its relationship with L-aspartic acid, which is present in Aginasa, but not in Leuginase. EcA2 shows a composition of monomers and oligomers up to tetramers, which is mostly not altered in the presence of L-Asp. The N-terminal loop of Leuginase, which is part of the active site is flexibly disordered, but gets ordered as in Aginasa in the presence os L-Asp, while L-Glu only does so to a limited extent. Ion-mobility spectrometry–mass spectrometry reveals two conformers for the monomeric EcA2, one of which can selectively bind to L-Asp and L-Glu. Aginasa has higher resistance to in vitro proteolysis than Leuginase, and this is directly related to the presence of L-Asp.Competing Interest StatementConflict of Interest: The authors have no financial conflicts of interest with the contents of this article. LMTRL is a participant in patent applications by the UFRJ on controlled release of peptides unrelated to the present work. The remaining authors have nothing to disclose.