Identification of Insulin-Activated Rab Proteins in Adipose Cells Using Bio-ATB-GTP Photolabeling Technique

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish
Title of host publicationGlucose Transport
Subtitle of host publicationMethods and Protocols
EditorsKarin Lindkvist-Petersson, Jesper S. Hansen
Place of PublicationNew York City, USA
PublisherHumana Press
Pages137-150
Number of pages14
ISBN (Electronic)978-1-4939-7507-5
ISBN (Print)978-1-4939-7506-8
DOIs
Publication statusE-pub ahead of print - 8 Dec 2017

Publication series

NameMethods in Molecular Biology
PublisherSpringer
Volume1713
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

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Keywords

  • Journal Article

Cite this

Koumanov, F., & Holman, G. D. (2017). Identification of Insulin-Activated Rab Proteins in Adipose Cells Using Bio-ATB-GTP Photolabeling Technique. In K. Lindkvist-Petersson, & J. S. Hansen (Eds.), Glucose Transport: Methods and Protocols (pp. 137-150). (Methods in Molecular Biology; Vol. 1713). New York City, USA: Humana Press. https://doi.org/10.1007/978-1-4939-7507-5_11