Cloning, expression, purification and crystallization of a transcriptional regulatory protein (Rv3291c) from Mycobacterium tuberculosis H37Rv.
Rv3291c, the translational product of the Mycobacterium tuberculosis Rv3291c gene, is an 18 kDa protein. It is a putative transcriptional regulatory protein belonging to the leucine-responsive regulatory protein/asparagine synthase C (Lrp/AsnC) family, which are proteins that have been identified in archaea and bacteria. Rv3291c probably plays a significant role during the persistent/latent phase of M. tuberculosis, as supported by its up-regulation several-fold during this stage. Orthorhombic crystals of recombinant Rv3291c have been grown from trisodium citrate dihydrate-buffered solutions containing monoammonium dihydrogen phosphate. Diffraction data extending to 2.7 A have been collected from a single crystal with unit-cell parameters a = 99.6, b = 100.7, c = 100.6 A. Assuming an octamer in the asymmetric unit results in a Matthews coefficient (VM) of 1.75 A3 Da(-1), corresponding to a solvent content of about 30%.
Shrivastava
Tripti
T
Molecular and Structural Biology Division, Central Drug Research Institute, PO Box 173, Chattar Manzil Palace, Mahatma Gandhi Marg, Lucknow 226001 (U.P.), India.
Kumar
Sandeep
S
Ramachandran
Ravishankar
R
D000595:Amino Acid Sequence; D002951:Citrates / Q000494:pharmacology; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D013058:Mass Spectrometry; D015233:Models, Statistical; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000378:metabolism*; D010710:Phosphates / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D000077559:Sodium Citrate; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013696:Temperature; D014157:Transcription Factors / Q000737:chemistry* / Q000235:genetics; D014158:Transcription, Genetic; D015854:Up-Regulation; D014961:X-Ray Diffraction
D002951:Citrates; D010710:Phosphates; D011994:Recombinant Proteins; C498065:Rv3291C protein, Mycobacterium tuberculosis; D014157:Transcription Factors; D000077559:Sodium Citrate