FEBS Letters | |
The Pro117 to glycine mutation of staphylococcal nuclease simplifies the unfolding—folding kinetics | |
Yamamoto, Kaori1  Ishihara, Tsuyoshi1  Kuwajima, Kunihiro1  Sugai, Shintaro1  Okayama, Naoki1  | |
[1] Department of Polymer Science, Faculty of Science, Hokkaido University, Kita-ku, Sapporo, Hokkaido 060, Japan | |
关键词: Staphylococcal nuclease; Folding mechanism; Proline isomerization; Stopped-flow technique; Circular dichroism; Site-directed mutagenesis; SNase; Staphylococcal nuclease; CD; circular dichroism; pdTp; thymidine 3′; 5′-diphosphate; NMR; nuclear magnetic resonance; Nc and Nt; two native species with the cis and trans forms; respectively; of the Lys116-Pro117 peptide bond; P117G; the Pro117 to glycine mutant; IPTG; isopropyl-β-D-thiogalactopyranoside; EGTA; [ethylenebis(oxyethylenenitrilo)]tetraacetic acid; | |
DOI : 10.1016/0014-5793(91)81243-2 | |
学科分类:生物化学/生物物理 | |
来源: John Wiley & Sons Ltd. | |
【 摘 要 】
Kinetics of unfolding and refolding of a staphylococcal nuclease mutant, in which Pro117 is replaced by glycine, have been investigated by stopped-flow circular dichroism, and the results are compared with those for the wild-type protein. In contrast to the biphasic unfolding of the wild-type nuclease, the unfolding of the mutant is represented by a single-phase reaction, indicating that the biphasic unfolding for the wild-type protein is caused by cis-trans isomerization about the prolyl peptide bond in the native state. The proline mutation also simplifies the kinetic refolding. Importance of the results in elucidating the folding mechanism is discussed.
【 授权许可】
Unknown
【 预 览 】
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