title: The Mycobacterium tuberculosis desaturase DesA1 (Rv0824c) is a Ca2+binding protein.
abstract: The hallmark feature of Mycobacterium tuberculosis (M.tb) the causative agent of human tuberculosis, is its complex lipid rich cell wall comprised primarily of mycolic acids, long chain fatty acids that play a key role in structural stability and permeability of the cell wall. In addition, they are involved in inhibiting phagosome-lysosome fusion and aid in granuloma formation during the pathogenic process. M.tb DesA1 is an essential acyl-acyl carrier protein desaturase predicted to catalyze the introduction of position specific double bonds during the biosynthesis of mycolic acids. This protein is one among three annotated desaturases (DesA1-3) in the M.tb genome but is unique in containing a βγ-crystallin Greek key signature motif, a well-characterized fold known to mediate Ca2+binding in both prokaryotic and eukaryotic organisms. Using Isothermal Titration Calorimetry and45CaCl2overlay, we demonstrate that Ca2+binds to DesA1. Spectroscopic measurements suggested that this binding induces changes in protein conformation but does not lead to significant alterations in the secondary structure of the protein, a feature common to several βγ-crystallins. An M. smegmatis strain over-expressing M.tb desA1 showed a Ca2+dependent variation in surface phenotype, revealing a functional role for Ca2+in DesA1 activity. This study represents the first identification of a Ca2+binding βγ-crystallin in M.tb, emphasizing the implicit role of Ca2+in the pathogenesis of M.tb.
authors.affiliation: CSIR - Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad 500007, India. Electronic address: veenachaitanya@ccmb.res.in.
authors.forename: Veena C
authors.initials: VC
authors.lastname: Yeruva
authors.affiliation: CSIR - Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad 500007, India.
authors.forename: Mamata
authors.initials: M
authors.lastname: Savanagouder
authors.affiliation: CSIR - Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad 500007, India.
authors.forename: Radhika
authors.initials: R
authors.lastname: Khandelwal
authors.affiliation: CSIR - Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad 500007, India.
authors.forename: Apoorva
authors.initials: A
authors.lastname: Kulkarni
authors.affiliation: CSIR - Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad 500007, India.
authors.forename: Yogendra
authors.initials: Y
authors.lastname: Sharma
authors.affiliation: CSIR - Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad 500007, India. Electronic address: raghu@ccmb.res.in.
authors.forename: Tirumalai R
authors.initials: TR
authors.lastname: Raghunand
mesh_terms: D000942:Antigens, Bacterial / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D002151:Calorimetry; D002942:Circular Dichroism; D044943:Fatty Acid Desaturases / Q000737:chemistry / Q000378:metabolism; D020102:Mycobacterium smegmatis / Q000235:genetics / Q000378:metabolism; D009169:Mycobacterium tuberculosis / Q000737:chemistry; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013050:Spectrometry, Fluorescence
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D002135:Calcium-Binding Proteins; D011994:Recombinant Proteins; C106712:desA1 protein, Mycobacterium tuberculosis; D044943:Fatty Acid Desaturases; D002118:Calcium
keywords: Ca(2+) binding protein; Desaturase; Mycobacterium tuberculosis; βɣ-Crystallin