- 著者
- Shingo Ueno Tatsunori Hirai Shusuke Sato Manish Biyani Hiromi Kuramochi Ryo Iizuka Takanori Akagi Takashi Funatsu Takanori Ichiki
- 出版者
- The Society of Photopolymer Science and Technology (SPST)
- 雑誌
- Journal of Photopolymer Science and Technology (ISSN:09149244)
- 巻号頁・発行日
- vol.28, no.5, pp.719-725, 2015-06-22 (Released:2015-10-20)
- 参考文献数
- 12
- 被引用文献数
- 1 4
The improved catalytic activity of enzymes is required in various fields. Enzymes have conventionally been improved by the screening of bacteria possessing mutant enzymes. However, the screening conditions are limited since screening requires the growth of bacteria. Here, we report the development of a protein microarray for the analysis of enzymatic activity. A his-tagged enzyme is synthesized in situ and immobilized on the microarray, which is composed of microreactors with a diameter and depth of 4 μm and a density of 1.0 x 106 reactors/cm2. β-glucosidase, synthesized in situ using a cell-free synthesis system, was immobilized on the microreactor array chip and its catalytic activity was observed. This enzyme-immobilized microarray is expected to enable the rapid and quantitative screening of enzymes.
- 著者
- Ryo Iizuka Takashi Funatsu
- 出版者
- 一般社団法人 日本生物物理学会
- 雑誌
- Biophysics and Physicobiology (ISSN:21894779)
- 巻号頁・発行日
- vol.13, pp.63-69, 2016 (Released:2016-04-22)
- 参考文献数
- 63
- 被引用文献数
- 1 9
The Escherichia coli chaperonin GroEL is an essential molecular chaperone that mediates protein folding in association with its cofactor, GroES. It is widely accepted that GroEL alternates the GroES-sealed folding-active rings during the reaction cycle. In other words, an asymmetric GroEL–GroES complex is formed during the cycle, whereas a symmetric GroEL–(GroES)2 complex is not formed. However, this conventional view has been challenged by the recent reports indicating that such symmetric complexes can be formed in the GroEL–GroES reaction cycle. In this review, we discuss the studies of the symmetric GroEL–(GroES)2 complex, focusing on the molecular mechanism underlying its formation. We also suggest that GroEL can be involved in two types of reaction cycles (asymmetric or symmetric) and the type of cycle used depends on the concentration of non-native substrate proteins.