TY - JOUR
T1 - Molecular basis of the functional divergence of fatty Acyl-AMP ligase biosynthetic enzymes of mycobacterium tuberculosis
AU - Goyal, Aneesh
AU - Verma, Priyanka
AU - Anandhakrishnan, Madhankumar
AU - Gokhale, Rajesh S.
AU - Sankaranarayanan, Rajan
N1 - Funding Information:
R.S. and R.S.G. acknowledge funding from Swarnajayanti Fellowship of Department of Science and Technology, India , and partial funding from European Union's Seventh Framework Programme . R.S. thanks support from National Bioscience Award of Department of Biotechnology, India . R.S.G. was supported by the HHMI International Investigator program . None of the authors have competing financial interests related to this work.
PY - 2012/2/17
Y1 - 2012/2/17
N2 - Activation of fatty acids as acyl-adenylates by fatty acyl-AMP ligases (FAALs) in Mycobacterium tuberculosis is a variant of a classical theme that involves formation of acyl-CoA (coenzyme A) by fatty acyl-CoA ligases (FACLs). Here, we show that FAALs and FACLs possess similar structural fold and substrate specificity determinants, and the key difference is the absence of a unique insertion sequence in FACL13 structure. A systematic analysis shows a conserved hydrophobic anchorage of the insertion motif across several FAALs. Strikingly, mutagenesis of two phenylalanine residues, which are part of the anchorage, to alanine converts FAAL32 to FACL32. This insertion-based in silico analysis suggests the presence of FAAL homologues in several other non-mycobacterial genomes including eukaryotes. The work presented here establishes an elegant mechanism wherein an insertion sequence drives the functional divergence of FAALs from canonical FACLs.
AB - Activation of fatty acids as acyl-adenylates by fatty acyl-AMP ligases (FAALs) in Mycobacterium tuberculosis is a variant of a classical theme that involves formation of acyl-CoA (coenzyme A) by fatty acyl-CoA ligases (FACLs). Here, we show that FAALs and FACLs possess similar structural fold and substrate specificity determinants, and the key difference is the absence of a unique insertion sequence in FACL13 structure. A systematic analysis shows a conserved hydrophobic anchorage of the insertion motif across several FAALs. Strikingly, mutagenesis of two phenylalanine residues, which are part of the anchorage, to alanine converts FAAL32 to FACL32. This insertion-based in silico analysis suggests the presence of FAAL homologues in several other non-mycobacterial genomes including eukaryotes. The work presented here establishes an elegant mechanism wherein an insertion sequence drives the functional divergence of FAALs from canonical FACLs.
KW - acyl-adenylate enzymes
KW - crystal structure
KW - pathogenesis
KW - substrate specificity
KW - virulent lipid synthesis
UR - http://www.scopus.com/inward/record.url?scp=84856437600&partnerID=8YFLogxK
U2 - 10.1016/j.jmb.2011.12.031
DO - 10.1016/j.jmb.2011.12.031
M3 - Article
C2 - 22206988
AN - SCOPUS:84856437600
SN - 0022-2836
VL - 416
SP - 221
EP - 238
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 2
ER -