TAILIEUCHUNG - Báo cáo khoa học: Engineering of monomeric FK506-binding protein 22 with peptidyl prolyl cis-trans isomerase Importance of a V-shaped dimeric structure for binding to protein substrate

FK506-binding protein 22 (FKBP22) from the psychrotrophic bacterium Shewanellasp. SIB1 is a homodimeric protein with peptidyl prolylcis–trans isomerase (PPIase) (EC ) activity. Each monomer consists of 205 amino acid residues. | Engineering of monomeric FK506-binding protein 22 with peptidyl prolyl cis-trans isomerase Importance of a V-shaped dimeric structure for binding to protein substrate Cahyo Budiman1 Keisuke Bando1 Clement Angkawidjaja1 Yuichi Koga1 Kazufumi Takano1 2 and Shigenori Kanaya1 1 Department of Materialand Life Science Graduate Schoolof Engineering Osaka University Yamadaoka Suita Osaka Japan 2 CRESTO JST Yamadaoka Suita Osaka Japan Keywords FKBP22 homodimer peptidyl-prolyl cis-trans isomerase PPIase protein engineering substrate binding Correspondence S. Kanaya Department of Materialand Life Science Graduate School of Engineering Osaka University 2-1 Yamadaoka Suita Osaka 565-0871 Japan Tel Fax 81 6 6879 7938 E-mail kanaya@ Received 1 May 2009 revised 24 May 2009 accepted 28 May 2009 doi FK506-binding protein 22 FKBP22 from the psychrotrophic bacterium Shewanella sp. SIB1 is a homodimeric protein with peptidyl prolyl cis-trans isomerase PPIase EC activity. Each monomer consists of 205 amino acid residues. According to a tertiary model SIB1 FKBP22 assumes a V-shaped structure in which two monomers interact with each other at their N-termini. Each monomer consists of an N-terminal domain with a dimerization core and a C-terminal catalytic domain which are separated by a 40-residue-long a-helix. To clarify the role of this V-shaped structure we constructed a mutant protein in which the N-domain is tandemly repeated through a flexible linker. This protein termed NNC-FKBP22 is designed such that two repetitive N-domains are folded into a structure similar to that of the Shewanella sp. SIB1 FKBP22 wild-type protein WT . NNC-FKBP22 was overproduced in Escherichia coli in a His-tagged form purified and biochemically characterized. Gel-filtration chromatography and ultracentrifugation analyses indicate that NNC-FKBP22 exists as a monomer. Analysis of thermal denaturation using differential scanning calorimetry .

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