Contribution of the C-Terminal Region of a Group II Chaperonin to its Interaction with Prefoldin and Substrate Transfer. Issue 11 (5th June 2016)

• Record Type: Journal Article
• Authors: Zako, Tamotsu; Sahlan, Muhamad; Fujii, Sayaka; Yamamoto, Yohei Y.; Tai, Phan The; Sakai, Kotaro; Maeda, Mizuo; Yohda, Masafumi
• Is Part Of: Journal of molecular biology. Volume 428:Issue 11(2016:Jun. 05)
• Page Start: 2405
• Page End: 2417
• Publication Date: 2016-06-05
• Subjects: CPN chaperonin -- T.KS-1 hyperthermophilic archaeon Thermococcus sp. strain KS-1 -- PFD prefoldin -- PFDα1β1 prefoldin of Thermococcus sp. strain KS-1 composed of α1 and β1 subunits -- PFDα2β2 prefoldin of Thermococcus sp. strain KS-1 composed of α2 and β2 subunits -- CS citrate synthase from porcine heart -- GFP green fluorescent protein -- SPR surface plasmon resonance -- PAGE polyacrylamide gel electrophoresis -- CPNβ T.KS-1 chaperonin β homo-oligomer -- CPNβTc1 T.KS-1 chaperonin β mutant with the truncation of 1 amino acid from the C-terminus -- CPNβTc2 T.KS-1 chaperonin β mutant with the truncation of 2 amino acids from the C-terminus -- CPNβTc6 T.KS-1 chaperonin β truncation mutant with the truncation of 6 amino acids from the C-terminus -- IPMDH 3-isopropylmalate dehydrogenase from Thermus thermophilus HB8 -- dIPMDH denatured IPMDH -- Cy3-dIPMDH Cy3-labeled dIPMDH -- 488-PFDα1β1 AlexaFluor 488-labeled PFDα1β1 -- Cy5-CPNβWT Cy5-labeled CPNβW -- Cy5-CPNβTc6 Cy5-labeled CPNβTc6 -- Cy5-CPNβs Cy5-labeled CPNβs -- PFDα1tcβ1 prefoldin of Thermococcus sp. strain KS-1 composed of two α1 subunits with truncation from the C-terminus and four β1 subunits -- RU resonance unit -- TKM buffer 50 mM Tris–HCl (pH 7.5), 100 mM KCl and 25 mM MgCl2; Chaperone -- Chaperonin -- Prefoldin -- Protein interaction -- Protein folding; Molecular biology -- Periodicals; Biology -- Periodicals; Biochemistry -- Periodicals; Bacteriology -- Periodicals; Molecular Biology -- Periodicals; Biochemistry -- Periodicals; Biologie moléculaire -- Périodiques; Biologie -- Périodiques; Biochimie -- Périodiques; Moleculaire biologie; Biochemistry; Biology; Molecular biology; Periodicals; 572.805
• DOI: 10.1016/j.jmb.2016.04.006
• Languages: English
• ISSNs: 0022-2836

Abstract

Abstract: Prefoldin is a molecular chaperone that captures an unfolded protein substrate and transfers it to a group II chaperonin. Previous studies have shown that the interaction sites for prefoldin are located in the helical protrusions of group II chaperonins. However, it does not exclude the possibility of the existence of other interaction sites. In this study, we constructed C-terminal truncation mutants of a group II chaperonin and examined the effects of these mutations on the chaperone's function and interaction with prefoldin. Whereas the mutants with up to 6 aa truncation from the C-terminus retained more than 90% chaperone activities for protecting citrate synthase from thermal aggregation and refolding of green fluorescent protein and isopropylmalate dehydrogenase, the truncation mutants showed decreased affinities for prefoldin. Consequently, the truncation mutants showed reduced transfer efficiency of the denatured substrate protein from prefoldin and subsequent chaperonin-dependent refolding. The results clearly show that the C-terminal region of group II chaperonins contributes to their interactions with prefoldin, the transfer of the substrate protein from prefoldin and its refolding. Graphical Abstract: Highlights: Prefoldin (PFD) interacts with group II chaperonin (CPN) for substrate transfer. The flexible C-terminal region of CPN protrudes into the cavity of CPN. Truncation of the CPN C-terminal region decreases affinity with PFD. Truncation also decreases substrate transfer from PFD and folding by CPN. The C-terminal region of CPN is a newly found interaction site with PFD.