Biochemical characterization of Mycobacterium tuberculosis LexA and structural studies of its C-terminal segment

Chandran, AV and Srikalaivani, R and Paul, A and Vijayan, M (2019) Biochemical characterization of Mycobacterium tuberculosis LexA and structural studies of its C-terminal segment. In: Acta Crystallographica Section D: Structural Biology, 75 (1). pp. 41-55.

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Abstract

LexA is a protein that is involved in the SOS response. The protein from Mycobacterium tuberculosis and its mutants have been biochemically characterized and the structures of their catalytic segments have been determined. The protein is made up of an N-terminal segment, which includes the DNA-binding domain, and a C-terminal segment encompassing much of the catalytic domain. The two segments are defined by a cleavage site. Full-length LexA, the two segments, two point mutants involving changes in the active-site residues (S160A and K197A) and another mutant involving a change at the cleavage site (G126D) were cloned and purified. The wild-type protein autocleaves at basic pH, while the mutants do not. The wild-type and the mutant proteins dimerize and bind DNA with equal facility. The C-terminal segment also dimerizes, and it also shows a tendency to form tetramers. The C-terminal segment readily crystallized. The crystals obtained from attempts involving the full-length protein and its mutants contained only the C-terminal segment including the catalytic core and a few residues preceding it, in a dimeric or tetrameric form, indicating protein cleavage during the long period involved in crystal formation. Modes of tetramerization of the full-length protein similar to those observed for the catalytic core are feasible. A complex of M. tuberculosis LexA and the cognate SOS box could be modeled in which the mutual orientation of the two N-terminal domains differs from that in the Escherichia coli LexA–DNA complex. These results represent the first thorough characterization of M. tuberculosis LexA and provide definitive information on its structure and assembly. They also provide leads for further exploration of this important protein.

Item Type:	Journal Article
Publication:	Acta Crystallographica Section D: Structural Biology
Publisher:	Wiley-Blackwell
Additional Information:	The copyright for this article belongs to Wiley-Blackwell.
Keywords:	bacterial protein; LexA protein, Bacteria; mutant protein; serine proteinase, chemistry; genetics; Mycobacterium tuberculosis; protein domain; protein multimerization; sos response (genetics), Bacterial Proteins; Mutant Proteins; Mycobacterium tuberculosis; Protein Domains; Protein Multimerization; Serine Endopeptidases; SOS Response (Genetics)
Department/Centre:	Division of Biological Sciences > Molecular Biophysics Unit
Date Deposited:	15 Nov 2022 07:26
Last Modified:	15 Nov 2022 07:26
URI:	https://eprints.iisc.ac.in/id/eprint/78023

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