Solution NMR Approaches for Establishing Specificity of Weak Heterodimerization of Membrane Proteins
| Autor: | Charles R. Sanders, Tiandi Zhuang, Bing K. Jap |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Vesicle-associated membrane protein 8
Magnetic Resonance Spectroscopy Sequence Homology Amino Acid biology Chemistry Molecular Sequence Data Peripheral membrane protein Membrane Proteins General Chemistry Nuclear magnetic resonance spectroscopy Biochemistry Article Catalysis Transmembrane protein Colloid and Surface Chemistry Membrane Membrane protein Amyloid precursor protein biology.protein Humans Electrophoresis Polyacrylamide Gel Amino Acid Sequence Dimerization Integral membrane protein |
| Zdroj: | Journal of the American Chemical Society. 133:20571-20580 |
| ISSN: | 1520-5126 0002-7863 |
| DOI: | 10.1021/ja208972h |
| Popis: | Solution NMR provides a powerful approach for detecting complex formation involving weak to moderate intermolecular affinity. However, solution NMR has only rarely been used to detect complex formation between two membrane proteins in model membranes. The impact of specific binding on the NMR spectrum of a membrane protein can be difficult to distinguish from spectral changes that are induced by non-specific binding and/or by changes that arise from forced co-habitation of the two proteins in a single model membrane assembly. This is particularly the case when solubility limits make it impossible to complete a titration to the point of near-saturation of complex formation. In this work experiments are presented that provide the basis for establishing whether specific complex formation occurs between two membrane proteins under conditions where binding is not of high avidity. Application of these methods led to the conclusion that the membrane protein CD147 (also known as EMMPRIN or basigin) forms a specific heterodimeric complex in the membrane with the 99 residue transmembrane C-terminal fragment of the amyloid precursor protein (C99 or APP-βCTF), the latter being the immediate precursor of the amyloid-β polypeptides that are closely linked to the etiology of Alzheimer’s disease. |
| Databáze: | OpenAIRE |
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