Specific localization and quantification of biotin transport components in yeast by use of a biotin-conjugated, impermeant, electron-dense label

Edward A. Bayer, Ehud Skutelsky, T. Viswanatha, Meir Wilchek

Research output: Contribution to journalArticlepeer-review

Abstract

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 biotinderivatized, 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.

Original languageEnglish
Pages (from-to)23-29
Number of pages7
JournalMolecular and Cellular Biochemistry
Volume19
Issue number1
DOIs
StatePublished - Feb 1978
Externally publishedYes

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