Heat shock protein 60 of filarial parasite Brugia malayi: cDNA cloning, expression, purification and in silico modeling and analysis of its ATP binding site.
We report here cloning and expression of full length mitochondrial HSP60 gene of Brugia malayi adult worm (mtHSP60bm), purification of the gene product by affinity chromatography, its in silico 3D structure and the sequence homology of the protein with Escherichia coli GroEL/ES and human HSP60. The ATP binding pocket of human HSP60 and mtHSP60bm were analyzed and compared using in silico models. The distribution of HSP60 in different life-stages of the parasite was determined using antibodies raised against recombinant mtHSP60bm (rmtHSP60bm). mtHSP60bm was present in all life-stages of the parasite except third stage infective larvae, in which it could be induced by heat-shock, and showed high degree of homology with E. coli GroEL/ES. The ATP binding pocket of HSP60 in humans, E. coli and B. malayi were also found structurally conserved. This similarity between human and mtHSP60bm might be useful in understanding the host-parasite interactions. This is the first ever report on distribution, cloning, sequence homology and ATP binding site of mtHSP60bm.
Division of Parasitology, CSIR-Central Drug Research Institute, Chattar Manzil Palace, Lucknow 226001, India.
R C
RC
Misra
A K
AK
Verma
S K
SK
Verma
V
V
Kumar
W A
WA
Siddiqui
M I
MI
Siddiqi
P K
PK
Murthy
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000330:Aedes; D000818:Animals; D001665:Binding Sites; D017178:Brugia malayi / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D018834:Chaperonin 60 / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D018509:DNA, Helminth / Q000235:genetics; D005260:Female; D018507:Gene Expression Regulation, Developmental; D005849:Gerbillinae; D006790:Host-Parasite Interactions; D006801:Humans; D007114:Immunization; D008297:Male; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D051189:Murinae; D018510:RNA, Helminth / Q000235:genetics / Q000302:isolation & purification; D017385:Sequence Homology
D018834:Chaperonin 60; D018076:DNA, Complementary; D018509:DNA, Helminth; D018510:RNA, Helminth; D000255:Adenosine Triphosphate