Chapter title |
Identification of Insulin-Activated Rab Proteins in Adipose Cells Using Bio-ATB-GTP Photolabeling Technique
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Chapter number | 11 |
Book title |
Glucose Transport
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Published in |
Methods in molecular biology, January 2018
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DOI | 10.1007/978-1-4939-7507-5_11 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7506-8, 978-1-4939-7507-5
|
Authors |
Francoise Koumanov, Geoffrey D. Holman, Koumanov, Francoise, Holman, Geoffrey D. |
Abstract |
We have recently developed a photolabeling method to identify GTP-loaded Rab proteins. The new biotinylated GTP analogue (Bio-ATB-GTP) binds to GTP-binding proteins and after a UV irradiation a covalent bond is formed between the protein and the photoreactive diazirine group on the photolabel. The tagged protein can then be isolated and detected using the classic biotin-streptavidin interaction. In this chapter, we describe the Bio-ATB-GTP photolabel and discuss the advantages of using this photolabeling approach to detect GTP-loaded Rab proteins compared to other existing methodologies. We also describe a step-by-step procedure for detecting the activated state of a Rab protein in primary rat adipocytes. |
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