Molecular Analysis of L-Asparaginases for Clarification of the Mechanism of Action and Optimization of Pharmacological Functions

Abstract

L-asparaginases (EC 3.5.1.1) are a family of enzymes that catalyze the hydrolysis of L-asparagine to L-aspartic acid and ammonia. These proteins with different biochemical, physicochemical and pharmacological properties are found in many organisms, including bacteria, fungi, algae, plants and mammals. To date, asparaginases from E. coli and Dickeya dadantii (formerly known as Erwinia chrysanthemi) are widely used in hematology for the treatment of lymphoblastic leukemias. However, their medical use is limited by side effects associated with the ability of these enzymes to hydrolyze L-glutamine, as well as the development of immune reactions. To solve these issues, gene-editing methods to introduce amino-acid substitutions of the enzyme are implemented. In this review, we focused on molecular analysis of the mechanism of enzyme action and to optimize the antitumor activity.

Molecular Mimicry of the Rheumatoid Arthritis-Related Immunodominant T-Cell Epitope within Type II Collagen (CII260-270) by the Bacterial L-Asparaginase

“…Bacterial L-asparaginase (L-ASNase, EC 3.5.1.1) shares amino acid sequence similarities (L-ASNase residues 68–78, IANVKGEQVVN) with CII260-270 [ 23 , 24 ]. This enzyme catalyzes the hydrolysis of L-asparagine to L-aspartate and ammonia and is absent in humans [ 24 , 25 , 26 ].…”

“…Bacterial L-asparaginase (L-ASNase, EC 3.5.1.1) shares amino acid sequence similarities (L-ASNase residues 68–78, IANVKGEQVVN) with CII260-270 [ 23 , 24 ]. This enzyme catalyzes the hydrolysis of L-asparagine to L-aspartate and ammonia and is absent in humans [ 24 , 25 , 26 ]. Many infectious pathogens suggested as major RA-triggering candidates ( Escherichia coli , Klebsiella pneumoniae , representatives of the genera Mycobacterium , Proteus , Streptococcus ) have been reported to produce L-asparaginase [ 25 ].…”

Electroenzymatic Model System for the Determination of Catalytic Activity of Erwinia carotovora L-Asparaginase

“…We assumed that amino acids with the ability for electrochemical oxidation, such as Tyr, Trp, His, Cys, Cys-Cys, and Met [19][20][21][22][23][24][25][26][27], are responsible for the response to substrate interactions. Tyr29 is a crucial amino acid in the active site of bacterial L-asparaginases and Tyr29 is the most important candidate for the main role in electrochemical registration of enzyme-substrate interactions in ASNase from Erwinia carotovora [31,47]. Tyr29 of L-asparaginase from Erwinia carotovora is involved in substrate binding and release of reaction product [31].…”

“…Tyr29 is a crucial amino acid in the active site of bacterial L-asparaginases and Tyr29 is the most important candidate for the main role in electrochemical registration of enzyme-substrate interactions in ASNase from Erwinia carotovora [31,47]. Tyr29 of L-asparaginase from Erwinia carotovora is involved in substrate binding and release of reaction product [31].…”

“…As shown earlier, Thr12, Tyr25, Thr89, Asp90, and Lys162 of ASNase participate in catalysis and substrate binding [29][30][31][32][33][34][35][36][37][38]. These amino acids are conserved for all bacterial asparaginases [29,[34][35][36][37][38][39][40][41][42].…”