| Content Provider |
World Health Organization (WHO)-Global Index Medicus |
| Author |
Nokhrin, Sergiy Barry, Amanda N. Yang, Haojun Dmitriev, Oleg Y. Markley, John L. Yu, Corey H. Huang, Yiping Hassanzadeh-ghassabeh, Gholamreza Tonelli, Marco Muyldermans, Serge Lutsenko, Svetlana |
| Description |
Author Affiliation: Huang Y ( From the Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.); Nokhrin S ( the Department of Biochemistry, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5E5, Canada.); Hassanzadeh-Ghassabeh G ( the Vrije Universiteit Brussel, Structural Biology Research Center, and Nanobody Service Facility, VIB, 1050 Brussels, Belgium, and.); Yu CH ( the Department of Biochemistry, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5E5, Canada.); Yang H ( From the Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.); Barry AN ( From the Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.); Tonelli M ( the Department of Biochemistry, National Magnetic Resonance Facility at Madison, University of Wisconsin-Madison, Madison, Wisconsin 53706.); Markley JL ( the Department of Biochemistry, National Magnetic Resonance Facility at Madison, University of Wisconsin-Madison, Madison, Wisconsin 53706.); Muyldermans S ( the Vrije Universiteit Brussel, Structural Biology Research Center, and.); Dmitriev OY ( the Department of Biochemistry, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5E5, Canada, Oleg.Dmitriev@usask.ca.); Lutsenko S ( From the Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, Lutsenko@jhmi.edu.) |
| Abstract |
The biologically and clinically important membrane transporters are challenging proteins to study because of their low level of expression, multidomain structure, and complex molecular dynamics that underlies their activity. ATP7B is a copper transporter that traffics between the intracellular compartments in response to copper elevation. The N-terminal domain of ATP7B (N-ATP7B) is involved in binding copper, but the role of this domain in trafficking is controversial. To clarify the role of N-ATP7B, we generated nanobodies that interact with ATP7B in vitro and in cells. In solution NMR studies, nanobodies revealed the spatial organization of N-ATP7B by detecting transient functionally relevant interactions between metal-binding domains 1-3. Modulation of these interactions by nanobodies in cells enhanced relocalization of the endogenous ATP7B toward the plasma membrane linking molecular and cellular dynamics of the transporter. Stimulation of ATP7B trafficking by nanobodies in the absence of elevated copper provides direct evidence for the important role of N-ATP7B structural dynamics in regulation of ATP7B localization in a cell. |
| ISSN |
00219258 |
| e-ISSN |
1083351X |
| Journal |
Journal of Biological Chemistry |
| Issue Number |
47 |
| Volume Number |
289 |
| Language |
English |
| Publisher |
American Society for Biochemistry and Molecular Biology (United States) |
| Publisher Date |
2014-11-21 |
| Publisher Place |
United States |
| Access Restriction |
Open |
| Subject Keyword |
Adenosine Triphosphatases Metabolism Cation Transport Proteins Copper Single-Domain Antibodies Chemistry Genetics Amino Acid Sequence Animals Binding Sites Blotting, Western Camelids, New World Cell Membrane HEK293 Cells Luminescent Proteins Magnetic Resonance Spectroscopy Microscopy, Confocal Models, Molecular Molecular Sequence Data Protein Binding Protein Structure, Tertiary Protein Transport Sequence Homology, Amino Acid Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Biochemistry Molecular Biology |
| Content Type |
Text |
| Resource Type |
Article |
| Subject |
Cell Biology Biochemistry Molecular Biology |
