New Insights Into the Function of Flavohemoglobin in Mycobacterium tuberculosis: Role as a NADPH-Dependent Disulfide Reductase and D-Lactate-Dependent Mycothione Reductase

被引:4
|
作者
Thakur, Naveen [1 ]
Sharma, Amar Nath [1 ]
Hade, Mangesh Dattu [1 ]
Chhaya, Ajay [2 ]
Kumar, Ashwani [1 ]
Jolly, Ravinder Singh [1 ]
Dikshit, Kanak L. [1 ,2 ]
机构
[1] CSIR, Inst Microbial Technol, Chandigarh, India
[2] Panjab Univ, Dept Biotechnol, Chandigarh, India
关键词
flavohemoglobin; Mycobacterium tuberculosis; FAD; D-lactate; thioredoxin reductase; oxidative stress; TRUNCATED HEMOGLOBIN; THIOREDOXIN; HBN; PEROXIDASE; EXPRESSION; RESISTANCE; SMEGMATIS; THIOL;
D O I
10.3389/fcimb.2021.796727
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Mycobacterium tuberculosis (Mtb) produces an unconventional flavohemoglobin (MtbFHb) that carries a FAD-binding site similar to D-lactate dehydrogenases (D-LDH) and oxidizes D-lactate into pyruvate. The molecular mechanism by which MtbFHb functions in Mtb remains unknown. We discovered that the D-LDH-type FAD-binding site in MtbFHb overlaps with another FAD-binding motif similar to thioredoxin reductases and reduces DTNB in the presence of NADPH similar to trxB of Mtb. These results suggested that MtbFHb is functioning as a disulfide oxidoreductase. Interestingly, D-lactate created a conformational change in MtbFHb and attenuated its ability to oxidize NADPH. Mass spectroscopy demonstrated that MtbFHb reduces des-myo-inositol mycothiol in the presence of D-lactate unlike NADPH, indicating that D-lactate changes the specificity of MtbFHb from di-thiol to di-mycothiol. When M. smegmatis carrying deletion in the fhbII gene (encoding a homolog of MtbFHb) was complemented with the fhb gene of Mtb, it exhibited four- to fivefold reductions in lipid peroxidation and significant enhancement in the cell survival under oxidative stress. These results were corroborated by reduced lipid peroxidation and enhanced cell survival of wild-type M. smegmatis after overexpression of the fhb gene of Mtb. Since D-lactate is a by-product of lipid peroxidation and MtbFHb is a membrane-associated protein, D-lactate-mediated reduction of mycothiol disulfide by MtbFHb may uniquely equip Mtb to relieve the toxicity of D-lactate accumulation and protect the cell from oxidative damage, simultaneously balancing the redox environment under oxidative stress that may be vital for the pathogenesis of Mtb.
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页数:15
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