Enzymatic activities of several K108 mutants of ribonuclease (RNase) Rh isolated from Rhizopus niveus.
We previously investigated the role of the Lys108 residue of ribonuclease (RNase) Rh from Rhizopus niveus, and suggested that Lys108 probably acts to stabilize the pentacovalent intermediate, and that an Arg residue could replace the role of Lys108. In RNase Le2 from Lentinus edodes, a homologous enzyme of RNase Rh, Lys108 is replaced by Thr. In this paper, the enzymatic properties of a K108T mutant and its analogous enzyme, K108S, were investigated to determine the effect of Thr and its analog, Ser at the 108th position on enzyme activity. The enzymatic properties of these mutant enzymes were compared with those of other mutant enzymes at this position (K108M, K108A, K108L). The results showed that Thr and Ser could replace Lys108 but resulted in only 2-20% of the activity of the native enzyme depending on the substrates used.
Ohgi
K
K
Department of Microbiology, Hoshi College of Pharmacy, Tokyo, Japan.
Iwama
M
M
Ogawa
Y
Y
Hagiwara
C
C
Ono
E
E
Kawaguchi
R
R
Kanazawa
C
C
Irie
M
M
D000595:Amino Acid Sequence; D008239:Lysine; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D012233:Rhizopus / Q000201:enzymology*; D012260:Ribonucleases / Q000737:chemistry / Q000378:metabolism*; D013329:Structure-Activity Relationship
D012260:Ribonucleases; D008239:Lysine