caa a22 4500 397556829 CHVBK 20180308164804.0 cr unu---uuuuu 161202e199608 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021406 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021406 Dimerization Characteristics of the 94-kDa Glucose-Regulated Protein [Elektronische Daten] [Takayuki Nemoto, Tomonori Matsusaka, Minoru Ota, Takashi Takagi, David B. Collinge, Haidee Walther-Larsen] The 94-kDa glucose-regulated protein (GRP94) is a member of the 90-kDa heat-shock protein (HSP90) family. In this study, we expressed the barley (Hordeum vulgare L.) GRP94 and the a isoform of human HSP90 (HSP90α) in Escherichia coli and compared their dimer-forming abilities. Native polyacrylamide gel electrophoresis revealed that GRP94 (amino acids 69-809) and the full-length form of HSP90α existed in the dimeric state. The C-terminal 326 amino acids of GRP94 or the C-terminal 200 amino acids of HSP90α were sufficient for the dimerization. Limited proteolysis of the C-terminal half of GRP94 with thrombin revealed a 16-kDa fragment, which was derived from the C-terminus of GRP94 through the cleavage of either the Arg710-His711 or the Arg735-Leu736 bond. These cleavage sites were nearly, if not completely, equivalent to the proteolyzed region of HSP90a. Their structural similarity prompted us to investigate, by use of a coexpression system, the possibility that the two proteins form a heterodimeric complex. A two-step affinity chromatography that specifically trapped only the complex revealed that the C-terminal 200 amino acids of HSP90α and the C-terminal 326 amino acids of GRP94 associated with HSP90α and GRP94, respectively. However, the C-terminal 326 amino acids of GRP94 failed to form a complex with HSP90α. In conclusion, these results indicate the similarity of the general dimeric conformation of the two HSP90 family member proteins, but show that the similarity is not sufficient to allow heterodimer formation. © 1996 BY THE JOURNAL OF BIOCHEMISTRY Regular Papers nationallicence amino acid sequence nationallicence coexpression system nationallicence dimerization nationallicence 94-kDa glucose-regulated protein nationallicence 90-kDa heat-shock protein nationallicence Nemoto Takayuki Department of Biochemistry, Iwate Medical University School of Dentistry 19-1 Uchimaru, Morioka, Iwate 020 aut Matsusaka Tomonori Department of Biochemistry, Iwate Medical University School of Dentistry 19-1 Uchimaru, Morioka, Iwate 020 aut Ota Minoru Department of Biochemistry, Iwate Medical University School of Dentistry 19-1 Uchimaru, Morioka, Iwate 020 aut Takagi Takashi Biological Institute, Graduate School of Science, Tohoku University Sendai 980-77 aut Collinge David B. Section of Plant Pathology, Department of Plant Biology, The Royal Veterinary and Agricultural University, Thorvaldsenvej 40, DK-1871 Frederiksberg C, Copenhagen, Denmark aut Walther-Larsen Haidee Section of Plant Pathology, Department of Plant Biology, The Royal Veterinary and Agricultural University, Thorvaldsenvej 40, DK-1871 Frederiksberg C, Copenhagen, Denmark aut The Journal of Biochemistry Oxford University Press 120/2(1996-08), 249-256 0021-924X 120:2<249 1996 120 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021406 text/html Onlinezugriff via DOI 1 other jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021406 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Nemoto Takayuki Department of Biochemistry, Iwate Medical University School of Dentistry 19-1 Uchimaru, Morioka, Iwate 020 aut NATIONALLICENCE 700 1- Matsusaka Tomonori Department of Biochemistry, Iwate Medical University School of Dentistry 19-1 Uchimaru, Morioka, Iwate 020 aut NATIONALLICENCE 700 1- Ota Minoru Department of Biochemistry, Iwate Medical University School of Dentistry 19-1 Uchimaru, Morioka, Iwate 020 aut NATIONALLICENCE 700 1- Takagi Takashi Biological Institute, Graduate School of Science, Tohoku University Sendai 980-77 aut NATIONALLICENCE 700 1- Collinge David B. Section of Plant Pathology, Department of Plant Biology, The Royal Veterinary and Agricultural University, Thorvaldsenvej 40, DK-1871 Frederiksberg C, Copenhagen, Denmark aut NATIONALLICENCE 700 1- Walther-Larsen Haidee Section of Plant Pathology, Department of Plant Biology, The Royal Veterinary and Agricultural University, Thorvaldsenvej 40, DK-1871 Frederiksberg C, Copenhagen, Denmark aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 120/2(1996-08), 249-256 0021-924X 120:2<249 1996 120 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford