Mitogenic factor secreted by Streptococcus pyogenes is a heat-stable nuclease requiring His122 for activity.
The gene encoding a mitogenic factor, termed MF, was cloned from Streptococcus pyogenes and the recombinant MF was overexpressed in Escherichia coli. Both the natural and recombinant MF had heat-resistant nuclease activity. The nuclease activity of MF was characterized using the recombinant protein. MF showed endonuclease activity, digesting ssDNA, dsDNA and tRNA. The optimal pH for the DNase activity of MF was 9.5. The DNase activity was enhanced approximately tenfold by the simultaneous presence of two divalent cations, Mg2+ and Ca2+, compared to either alone and was inhibited by EDTA or NaCl. The heat stability of MF was biphasic; the DNase activity was heat-stable from 0 to 50 degrees C over 80 degrees C but very unstable at around 60 degrees C. DNA digested by MF possessed 5'-phosphorylated and 3'-hydroxylated termini, identical to those obtained by digestion of DNA by pancreatic deoxyribonuclease I. A mutant clone revealed that His122 was a residue essential to the nuclease activity.
Iwasaki
Makoto
M
Shionogi Institute for Medical Science, 2-5-1 Mishima, Settsu, Osaka 566, Japan.
Igarashi
Hisanaga
H
Shionogi Institute for Medical Science, 2-5-1 Mishima, Settsu, Osaka 566, Japan.
Yutsudo
Takashi
T
Discovery Research laboratory I, Shionogi & Co., Ltd, 3-1-1 Futaba-cho, Toyonaka-shi, Osaka 561, Japan.
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D003851:Deoxyribonucleases / Q000235:genetics* / Q000378:metabolism; D004789:Enzyme Activation / Q000235:genetics; D006639:Histidine / Q000235:genetics; D006358:Hot Temperature; D008934:Mitogens / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013297:Streptococcus pyogenes / Q000235:genetics* / Q000378:metabolism
D001426:Bacterial Proteins; D008934:Mitogens; D011994:Recombinant Proteins; D006639:Histidine; D003851:Deoxyribonucleases