期刊
MOLECULAR BIOSYSTEMS
卷 6, 期 10, 页码 1829-1833出版社
ROYAL SOC CHEMISTRY
DOI: 10.1039/c005223j
关键词
-
资金
- NSF of China [20935005, 90813032, 20875092, 20905070]
- Ministry of Science and Technology of China [2008AA02Z206, 2010CB933502]
- Chinese Academy of Sciences
Rhodamine B piperazinoacetohydrazine (RBPH) is used as a bifunctional probe for the N-terminal specific modification of a thermophilic enzyme (T. lanuginosus xylanase), and the modification effect on the thermostability of the enzyme is investigated. The probe RBPH, bearing a spectroscopic unit of rhodamine B, a carbonyl-specific labeling unit of hydrazine and a linker of piperazine, not only has a stable always-on spectroscopic response, but also exists in a cationic form. These properties enable RBPH to serve as a bifunctional probe (simultaneous introduction of stable spectroscopic signal and positive charge) for the protein modification, and such an application has been successfully demonstrated on the N-terminal labeling of T. lanuginosus xylanase. A temperature-dependent inactivation study shows that the modification of T. lanuginosus xylanase by RBPH hardly changes its thermostability, in other words, a small change in electric charge of the N-terminal region caused by introducing one positive charge is not enough to alter the thermostability of the enzyme. This reveals a conservative property of the N-terminal domain for electric charge change, and such a property may result from the fact that the N-terminal domain of the enzyme already has 4 charged residues, which can produce strong electrostatic interactions, thereby making the domain quite stable.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据