期刊
JOURNAL OF BIOMOLECULAR STRUCTURE & DYNAMICS
卷 35, 期 5, 页码 1030-1041出版社
TAYLOR & FRANCIS INC
DOI: 10.1080/07391102.2016.1169219
关键词
Mycobacterium tuberculosis; isocitrate lyase; lysine succinylation; site-directed mutagenesis; crystal structure; antibiotics resistance
资金
- National Natural Science Foundation [81371851, 81511120001, 81071316, 81271882, 81301394, 81172806, 81471563]
- New Century Excellent Talents in Universities [NCET-11-0703]
- National Megaprojects for Key Infectious Diseases [2008ZX10003-006, 2008ZX10003-001]
- Southwest University [kb2010017, ky2011003]
- Fundamental Research Funds for the Central Universities [XDJK2011D006, XDJK2012D011, XDJK2012D007, XDJK2013D003, XDJK2014D040, XDJK2016D025]
- Graduate research and innovation project of graduate in Chongqing [CYS14044]
- Chongqing Municipal Committee of Education for postgraduates excellence program [YJG123104]
- Chongqing Municipal Committee of Education for undergraduates teaching reform program [2013JY201]
Lysine succinylation (Ksucc) is a newly identified protein posttranslational modification (PTM), which may play an important role in cellular physiology. However, the role of lysine succinylation in antibiotic resistance remains elusive. Isocitrate lyase (ICL) is crucial for broad-spectrum antibiotics tolerance in Mycobacterium tuberculosis (Mtb). We previously found that MtbICL (Rv0467) has at least three succinylated lysine residues, namely K189, K322, and K334.To explore the effect of succinylation on the activity of MtbICL, mutants' mimicry of the lysine succinylation were generated by site-directed mutagenesis. ICL-K189E mutant strain is more sensitive than the wild-type to rifampicin and streptomycin, but not isoniazid. For the in vitro activity of the purified isocitrate lyase, only K189E mutant showed significantly decreased activity. Crystal structure analysis showed that Lys189 Glu dramatically increased the pKa of Glu188 and decreased the pKa of Lys190, whereas had negligible effect on other residues within 5 angstrom as well as disruption of the electrostatic interaction between Lys189 and Glu182, which might prevent the closure of the active site loop and cause severe reduction of the enzyme activity. Considering the genetic, biochemical, and crystallographical evidences together, the succinylation of specific ICL residue can fine-tune the bacterial resistance to selected antibiotics. The decreased enzymatic activity resulting from the succinylation-changed electrostatic interaction might underlie this phenotype. This study provided the first insight into the link between lysine succinylation and antibiotic resistance.
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