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Structural characterization of a putative recombinant l-amino acid oxidase from Leptospira interrogans


Affiliations
1 Genomics and Central Research Laboratory, Department of Cell Biology and Molecular Genetics, Sri Devaraj Urs Academy of Higher Education and Research, Tamaka, Kolar 563 101, India, India
2 Computational Data Sciences, Indian Institute of Science, Bengaluru 560 012, India; Present address: National Institute for Plant Biotechnology, Indian Council of Agricultural Research, New Delhi 110 012, India, India
3 Molecular Biophysics Unit, Indian Institute of Science, Bengaluru 560 012, India, India
4 Computational Data Sciences, Indian Institute of Science, Bengaluru 560 012, India, India
5 Molecular Biophysics Unit, Indian Institute of Science, Bengaluru 560 012, India; Institute of Bioinformatics and Applied Biotechnology, Bengaluru 560 100, India, India
6 Genomics and Central Research Laboratory, Department of Cell Biology and Molecular Genetics, Sri Devaraj Urs Academy of Higher Education and Research, Tamaka, Kolar 563 101, India; Centre for Nanoscience and Engineering, Indian Institute of Science, Bengaluru 560 012, India, India
 

Amino acid oxidases (AOs) are flavin adenine dinucleotide (FAD)-dependent dimeric enzymes that stereo specifically catalyse the deamination of an a-amino acid leading to an a-keto acid. Putative Leptospira interrogans recombinant l-amino acid oxidase (Li-rLAO; lacking 20 residues corresponding to the N-terminal signal sequence) was cloned, expressed, purified, and its three-dimensional structure was determined by X-ray crystallography at a resolution of 1.8 Å. The active site could be easily identified by the presence of electron density corresponding to a non-covalently bound FAD in both protomers of the dimeric enzyme. Structural analysis of Li-rLAO revealed that its polypeptide fold is similar to those of the previously determined homologous structures as available in the Protein Data Bank. However, a substrate-binding residue found at the active site of other previously determined homologous structures was not conserved in Li-rLAO, suggesting that its specificity may differ from those of earlier reported structures. Not surprisingly, Li-rLAO showed no activity for most amino acids and amines; it exhibited a low activity only with l-arginine as the substrate. The catalytic properties of Li-rLAO could be rationalized in terms of its three-dimensional structure
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Abstract Views: 398

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  • Structural characterization of a putative recombinant l-amino acid oxidase from Leptospira interrogans

Abstract Views: 398  |  PDF Views: 168

Authors

D. Vaigundan
Genomics and Central Research Laboratory, Department of Cell Biology and Molecular Genetics, Sri Devaraj Urs Academy of Higher Education and Research, Tamaka, Kolar 563 101, India, India
I. Yuvaraj
Computational Data Sciences, Indian Institute of Science, Bengaluru 560 012, India; Present address: National Institute for Plant Biotechnology, Indian Council of Agricultural Research, New Delhi 110 012, India, India
P. Sunita
Molecular Biophysics Unit, Indian Institute of Science, Bengaluru 560 012, India, India
K. Sekar
Computational Data Sciences, Indian Institute of Science, Bengaluru 560 012, India, India
M. R. N. Murthy
Molecular Biophysics Unit, Indian Institute of Science, Bengaluru 560 012, India; Institute of Bioinformatics and Applied Biotechnology, Bengaluru 560 100, India, India
P. R. Krishnaswamy
Genomics and Central Research Laboratory, Department of Cell Biology and Molecular Genetics, Sri Devaraj Urs Academy of Higher Education and Research, Tamaka, Kolar 563 101, India; Centre for Nanoscience and Engineering, Indian Institute of Science, Bengaluru 560 012, India, India

Abstract


Amino acid oxidases (AOs) are flavin adenine dinucleotide (FAD)-dependent dimeric enzymes that stereo specifically catalyse the deamination of an a-amino acid leading to an a-keto acid. Putative Leptospira interrogans recombinant l-amino acid oxidase (Li-rLAO; lacking 20 residues corresponding to the N-terminal signal sequence) was cloned, expressed, purified, and its three-dimensional structure was determined by X-ray crystallography at a resolution of 1.8 Å. The active site could be easily identified by the presence of electron density corresponding to a non-covalently bound FAD in both protomers of the dimeric enzyme. Structural analysis of Li-rLAO revealed that its polypeptide fold is similar to those of the previously determined homologous structures as available in the Protein Data Bank. However, a substrate-binding residue found at the active site of other previously determined homologous structures was not conserved in Li-rLAO, suggesting that its specificity may differ from those of earlier reported structures. Not surprisingly, Li-rLAO showed no activity for most amino acids and amines; it exhibited a low activity only with l-arginine as the substrate. The catalytic properties of Li-rLAO could be rationalized in terms of its three-dimensional structure

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DOI: https://doi.org/10.18520/cs%2Fv123%2Fi7%2F895-906