Molecular characterization of GroES and GroEL homologues from Clostridium botulinum

• Published in: Journal of Protein Chemistry Vol. 22; no. 1; pp. 99 - 108
• Main Authors: Sagane, Yoshimasa, Hasegawa, Kimiko, Mutoh, Shingo, Kouguchi, Hirokazu, Suzuki, Tomonori, Sunagawa, Hiroyuki, Nakagawa, Tomoyuki, Kamaguchi, Arihide, Okasaki, Shinn, Nakayama, Kenji, Watanabe, Toshihiro, Oguma, Keiji, Ohyama, Tohru
• Format: Journal Article
• Language: English
• Published: United States Springer Nature B.V 01.01.2003
• Subjects: Amino Acid Sequence; Amino acids; Antibodies; Bacteria; Bacterial proteins; Base Sequence; Botulinum toxin type D; Chaperonin 10 - chemistry; Chaperonin 10 - genetics; Chaperonin 10 - isolation & purification; Chaperonin 60 - chemistry; Chaperonin 60 - genetics; Chaperonin 60 - isolation & purification; Clostridium botulinum; Clostridium botulinum - chemistry; Clostridium botulinum - genetics; Deoxyribonucleic acid; DNA; DNA, Bacterial - genetics; E coli; Genes, Bacterial - genetics; Genomics; GroEL gene; Homology; Immunoglobulins; Microbiology; Molecular biology; Molecular Sequence Data; Open reading frames; Operon - genetics; Proteins; Sequence Homology; Toxins
• ISSN: 0277-8033; 1572-3887; 1573-4943
• DOI: 10.1023/a:1023028113566

We report novel findings of significant amounts of 60- and 10-kDa proteins on SDS-PAGE in a culture supernatant of the Clostridium botulinum type D strain 4947 (D-4947). The N-terminal amino acid sequences of the purified proteins were closely related to those of other bacterial GroEL and GroES proteins, and both positively cross-reacted with Escherichia coli GroEL and GroES antibodies. Native GroEL homologue as an oligomeric complex is a weak ATPase whose activity is inhibited by the presence of GroES homologue. The 2634-bp groESL operon of D-4947 was isolated by PCR and sequenced. The sequence included two complete open reading frames (282 and 1629 bp), which were homologous to the groES and groEL gene family of bacterial proteins. Southern and Northern blot analyses indicate that the groESL operon is encoded on the genomic DNA of D-4947 as a single copy, and not on that of its specific toxin-converting phage.