Specific localization and quantification of biotin transport components in yeast by use of a biotin-conjugated, impermeant, electron-dense label
✍ Scribed by Edward A. Bayer; Ehud Skutelsky; T. Viswanatha; Meir Wilchek
- Publisher
- Springer
- Year
- 1978
- Tongue
- English
- Weight
- 527 KB
- Volume
- 19
- Category
- Article
- ISSN
- 0300-8177
No coin nor oath required. For personal study only.
✦ Synopsis
Two approaches are described for the localization and quantification of biotin transport components in yeast cells. One approach is based on tracing the fate of a radioactive affinity label for the biotin transport system, [14C]biotinyl-p-nitrophenyl ester (pBNP), through various stages of subcellular fractionations. A complementary method involves the use of a biotin-derivatized, impermeant, electron-dense, affinity-cytochemical label (ferritin-biotin conjugates) for subsequent visualization by electron microscopy. Values of approximately 8,000 and 4,000 sites/cell, respectively, were achieved by the two methods. Complicating factors, future perspectives and the relevance of the two methods to the isolation of transport components are discussed.