| Content Provider |
World Health Organization (WHO)-Global Index Medicus |
| Author |
Huang, Yun-tzu Pan, Rong-long Hsu, Shen-hsing Lo, Yueh-yu Sun, Yuh-ju Tseng, Fan-gang Pan, Yih-jiuan Hung, Cheng-chieh Liu, Tseng-huang Yang, Chih-wei |
| Description |
Author Affiliation: Hsu SH ( From the Department of Nephrology, Kidney Research Center, Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Taoyuan 33333.); Lo YY ( From the Department of Nephrology, Kidney Research Center, Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Taoyuan 33333.); Liu TH ( the Department of Life Science and Institute of Bioinformatics and Structural Biology, College of Life Science, and.); Pan YJ ( the Department of Life Science and Institute of Bioinformatics and Structural Biology, College of Life Science, and.); Huang YT ( the Department of Life Science and Institute of Bioinformatics and Structural Biology, College of Life Science, and.); Sun YJ ( the Department of Life Science and Institute of Bioinformatics and Structural Biology, College of Life Science, and.); Hung CC ( From the Department of Nephrology, Kidney Research Center, Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Taoyuan 33333.); Tseng FG ( Department of Engineering and System Science, College of Nuclear Science, National Tsing Hua University, Hsin Chu 30013, Taiwan.); Yang CW ( From the Department of Nephrology, Kidney Research Center, Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Taoyuan 33333, cwyang@ms1.hinet.net.); Pan RL ( the Department of Life Science and Institute of Bioinformatics and Structural Biology, College of Life Science, and rlpan@life.nthu.edu.tw.) |
| Abstract |
Single molecule atomic force microscopy (smAFM) was employed to unfold transmembrane domain interactions of a unique vacuolar H(+)-pyrophosphatase (EC 3.6.1.1) from Vigna radiata. H(+)-Pyrophosphatase is a membrane-embedded homodimeric protein containing a single type of polypeptide and links PPi hydrolysis to proton translocation. Each subunit consists of 16 transmembrane domains with both ends facing the lumen side. In this investigation, H(+)-pyrophosphatase was reconstituted into the lipid bilayer in the same orientation for efficient fishing out of the membrane by smAFM. The reconstituted H(+)-pyrophosphatase in the lipid bilayer showed an authentically dimeric structure, and the size of each monomer was â ¼4 nm in length, â ¼2 nm in width, and â ¼1 nm in protrusion height. Upon extracting the H(+)-pyrophosphatase out of the membrane, force-distance curves containing 10 peaks were obtained and assigned to distinct domains. In the presence of pyrophosphate, phosphate, and imidodiphosphate, the numbers of interaction curves were altered to 7, 8, and 10, respectively, concomitantly with significant modification in force strength. The substrate-binding residues were further replaced to verify these domain changes upon substrate binding. A working model is accordingly proposed to show the interactions between transmembrane domains of H(+)-pyrophosphatase in the presence and absence of substrate and its analog. |
| ISSN |
00219258 |
| e-ISSN |
1083351X |
| Journal |
Journal of Biological Chemistry |
| Issue Number |
2 |
| Volume Number |
290 |
| Language |
English |
| Publisher |
American Society for Biochemistry and Molecular Biology (United States) |
| Publisher Date |
2015-01-09 |
| Publisher Place |
United States |
| Access Restriction |
Open |
| Subject Keyword |
Inorganic Pyrophosphatase Chemistry Ultrastructure Ion Transport Vacuoles Enzymology Fabaceae Hydrolysis Metabolism Kinetics Lipid Bilayers Microscopy, Atomic Force Protein Structure, Tertiary Protons Substrate Specificity Research Support, Non-U.S. Gov't Biochemistry Molecular Biology |
| Content Type |
Text |
| Resource Type |
Article |
| Subject |
Cell Biology Biochemistry Molecular Biology |
