Thiol protection in membrane protein purifications: A study with phage holins
| Autor: | Douglas K. Struck, Jill S. Dewey, Ry Young |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2009 |
| Předmět: |
chemistry.chemical_classification
Vesicle-associated membrane protein 8 Lysis Chemistry Peripheral membrane protein Molecular Sequence Data Biophysics Membrane Proteins Cell Biology Biochemistry Article Viral Proteins 2 2'-Dipyridyl Membrane protein Holin Thiol Bacteriophages Amino Acid Sequence Cysteine Disulfides Molecular Biology Integral membrane protein |
| Popis: | The lambda holin, or S105, is a small cytoplasmic membrane protein that controls the timing of host lysis. Using thiol-specific reagents, we determined that the single cysteine residue within S105 was heterogeneously modified during membrane extraction and subsequent immobilized metal ion chromatography. Here we describe the use of a specific and reversible thiol reagent, 2,2′-dithiodipyridine, to generate purified protein with its cysteine residues in the native thiol state. The 2,2′-dithiodipyridine protection protocol was also successfully used for another unrelated holin, S 21 68, and should be generally useful for the purification of membrane proteins. |
| Databáze: | OpenAIRE |
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