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- Title
The role of Glu196 in the environment around the substrate binding site of leucine aminopeptidase from Streptomyces griseus
- Authors
Arima, Jiro; Uesugi, Yoshiko; Uraji, Misugi; Iwabuchi, Masaki; Hatanaka, Tadashi
- Abstract
Abstract: To investigate the role of Glu196 of leucine aminopeptidase from Streptomyces griseus (SGAP) in SGAP activation by calcium and substrate specificity, we constructed E196X SGAP by saturation mutagenesis. Most mutations led to the abrogation of SGAP activation by calcium, and substitution with Lys led to a marked increase in activity toward Asp-p-nitroanilide (pNA) and a decrease in that toward Lys-pNA. A similar result was obtained from the investigation using non-calcium-activated enzyme from Streptomyces septatus (SSAP). These results indicate that Glu196 of SGAP is associated with the environment around the substrate binding site besides its role in SGAP activation by calcium.
- Subjects
STREPTOMYCES griseus; LEUCINE aminopeptidase; CALCIUM; MUTAGENESIS
- Publication
FEBS Letters, 2006, Vol 580, Issue 3, p912
- ISSN
0014-5793
- Publication type
Article
- DOI
10.1016/j.febslet.2006.01.014