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

Francoise Koumanov; Geoffrey D Holman

doi:10.1007/978-1-4939-7507-5_11

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

Research output: Chapter or section in a book/report/conference proceeding › Chapter or section

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 language	English
Title of host publication	Glucose Transport
Subtitle of host publication	Methods and Protocols
Editors	Karin Lindkvist-Petersson, Jesper S. Hansen
Place of Publication	New York City, USA
Publisher	Humana Press
Pages	137-150
Number of pages	14
ISBN (Electronic)	978-1-4939-7507-5
ISBN (Print)	978-1-4939-7506-8
DOIs	https://doi.org/10.1007/978-1-4939-7507-5_11
Publication status	E-pub ahead of print - 8 Dec 2017

Publication series

Name	Methods in Molecular Biology
Publisher	Springer
Volume	1713
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Keywords

Journal Article

Access to Document

10.1007/978-1-4939-7507-5_11

Role of Rab3 in Peripheral Tissue Insulin Resistance
Koumanov, F. (PI), Betts, J. (CoI), Gonzalez, J. (CoI), Thompson, D. (CoI) & Whitley, P. (CoI)
MRC
1/12/16 → 31/07/20
Project: Research council
Elucidation of Final Stages in Coupling of Insulin Signalling to CLUT4 Translocation
Holman, G. (PI) & Koumanov, F. (CoI)
MRC
1/07/12 → 30/06/15
Project: Research council
Rab Proteins as Regulated Intermediates in Insulin-and Energy-Status Stimulated GLUT4 Trafficking
Holman, G. (PI) & Koumanov, F. (CoI)
Diabetes UK
1/07/09 → 30/06/11
Project: UK charity

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). Humana Press. Advance online publication. https://doi.org/10.1007/978-1-4939-7507-5_11

Identification of Insulin-Activated Rab Proteins in Adipose Cells Using Bio-ATB-GTP Photolabeling Technique. / Koumanov, Francoise ; Holman, Geoffrey D.
Glucose Transport: Methods and Protocols. ed. / Karin Lindkvist-Petersson; Jesper S. Hansen. New York City, USA: Humana Press, 2017. p. 137-150 (Methods in Molecular Biology; Vol. 1713).

Research output: Chapter or section in a book/report/conference proceeding › Chapter or section

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

Abstract

Publication series

Keywords

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Projects

Role of Rab3 in Peripheral Tissue Insulin Resistance

Elucidation of Final Stages in Coupling of Insulin Signalling to CLUT4 Translocation

Rab Proteins as Regulated Intermediates in Insulin-and Energy-Status Stimulated GLUT4 Trafficking

Cite this