Molecular characterization of GroES and GroEL homologues from Clostridium botulinum.
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.
Sagane
Yoshimasa
Y
Department of Food Science and Technology, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri 099-2493, Japan.
Hasegawa
Kimiko
K
Mutoh
Shingo
S
Kouguchi
Hirokazu
H
Suzuki
Tomonori
T
Sunagawa
Hiroyuki
H
Nakagawa
Tomoyuki
T
Kamaguchi
Arihide
A
Okasaki
Shinn
S
Nakayama
Kenji
K
Watanabe
Toshihiro
T
Oguma
Keiji
K
Ohyama
Tohru
T
D000595:Amino Acid Sequence; D001483:Base Sequence; D018835:Chaperonin 10 / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D018834:Chaperonin 60 / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D003014:Clostridium botulinum / Q000737:chemistry* / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics*; D005798:Genes, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D009876:Operon / Q000235:genetics; D017385:Sequence Homology
D018835:Chaperonin 10; D018834:Chaperonin 60; D004269:DNA, Bacterial