A mechanism and an evaluation of surface specific iodination by the chloramine-T procedure. |
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Authors: | R C Montelaro R R Rueckert |
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Institution: | Biophysics Laboratory of the Graduate School and Biochemistry Department of the College of Agricultural and Life Sciences, University of Wisconsin, Madison, Wisconsin 53706 U.S.A. |
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Abstract: | Both internal and external proteins in vesicular stomatitis virus were labeled when intact virions were iodinated with 50 μm iodide; however, only the surface proteins were labeled when the same procedure was carried out at low iodide concentrations (below 0.5 μm). This result together with similar observations reported earlier with another enveloped virus, Rous-associated virus-61 (RAV-61), suggest that viral envelopes provide a barrier to iodination by chloramine-T at low, but not at high, iodide concentrations. By monitoring the permeability of the RAV-61 envelope to successive iodinations and to iodination in the presence of chaotropic thiocyanate ions, it was shown that the permeability of the viral envelope was not altered at the higher concentrations of iodide. Further results support the hypothesis that iodination mediated by chloramine-T inolves two different iodinating species: (a) a membrane impermeable one, possibly “iodamine-T,” which predominates at low iodide concentrations, and (b) a membrane permeable species, possibly molecular iodine, which predominates at high concentrations of iodide. These results reinforce the proposal that the chloramine-T procedure is a useful method for specifically labeling surface proteins of lipid-enveloped structures. |
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Keywords: | To whom correspondence should be addressed Present address: Department of Surgery Tumor Virology Laboratory P O Box 2926 Duke University Medical Center Durham North Carolina 27710 |
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