Discovery - University of Dundee - Online Publications

Library & Learning Centre

Thr(649)Ala-AS160 knock-in mutation does not impair contraction/AICAR-induced glucose transport in mouse muscle

Standard

Thr(649)Ala-AS160 knock-in mutation does not impair contraction/AICAR-induced glucose transport in mouse muscle. / Ducommun, Serge; Wang, Hong Yu; Sakamoto, Kei; MacKintosh, Carol; Chen, Shuai.

In: American Journal of Physiology, Endocrinology and Metabolism, Vol. 302, No. 9, 05.2012, p. E1036-E1043.

Research output: Contribution to journalArticle

Harvard

Ducommun, S, Wang, HY, Sakamoto, K, MacKintosh, C & Chen, S 2012, 'Thr(649)Ala-AS160 knock-in mutation does not impair contraction/AICAR-induced glucose transport in mouse muscle' American Journal of Physiology, Endocrinology and Metabolism, vol 302, no. 9, pp. E1036-E1043.

APA

Ducommun, S., Wang, H. Y., Sakamoto, K., MacKintosh, C., & Chen, S. (2012). Thr(649)Ala-AS160 knock-in mutation does not impair contraction/AICAR-induced glucose transport in mouse muscle. American Journal of Physiology, Endocrinology and Metabolism, 302(9), E1036-E1043doi: 10.1152/ajpendo.00379.2011

Vancouver

Ducommun S, Wang HY, Sakamoto K, MacKintosh C, Chen S. Thr(649)Ala-AS160 knock-in mutation does not impair contraction/AICAR-induced glucose transport in mouse muscle. American Journal of Physiology, Endocrinology and Metabolism. 2012 May;302(9):E1036-E1043.

Author

Ducommun, Serge; Wang, Hong Yu; Sakamoto, Kei; MacKintosh, Carol; Chen, Shuai / Thr(649)Ala-AS160 knock-in mutation does not impair contraction/AICAR-induced glucose transport in mouse muscle.

In: American Journal of Physiology, Endocrinology and Metabolism, Vol. 302, No. 9, 05.2012, p. E1036-E1043.

Research output: Contribution to journalArticle

Bibtex - Download

@article{ac6136b09c644f4889e1e506c389ac7f,
title = "Thr(649)Ala-AS160 knock-in mutation does not impair contraction/AICAR-induced glucose transport in mouse muscle",
author = "Serge Ducommun and Wang, {Hong Yu} and Kei Sakamoto and Carol MacKintosh and Shuai Chen",
year = "2012",
volume = "302",
number = "9",
pages = "E1036--E1043",
journal = "American Journal of Physiology, Endocrinology and Metabolism",
issn = "0193-1849",

}

RIS (suitable for import to EndNote) - Download

TY - JOUR

T1 - Thr(649)Ala-AS160 knock-in mutation does not impair contraction/AICAR-induced glucose transport in mouse muscle

A1 - Ducommun,Serge

A1 - Wang,Hong Yu

A1 - Sakamoto,Kei

A1 - MacKintosh,Carol

A1 - Chen,Shuai

AU - Ducommun,Serge

AU - Wang,Hong Yu

AU - Sakamoto,Kei

AU - MacKintosh,Carol

AU - Chen,Shuai

PY - 2012/5

Y1 - 2012/5

N2 - <p>Ducommun S, Wang HY, Sakamoto K, MacKintosh C, Chen S. Thr(649)Ala-AS160 knock-in mutation does not impair contraction/AICAR-induced glucose transport in mouse muscle. Am J Physiol Endocrinol Metab 302: E1036-E1043, 2012. First published February 7, 2012; doi:10.1152/ajpendo.00379.2011.-AS160 and its closely related protein TBC1D1 have emerged as key mediators for both insulin-and contraction-stimulated muscle glucose uptake through regulating GLUT4 trafficking. Insulin increases AS160 phosphorylation at multiple Akt/PKB consensus sites, including Thr(649), and promotes its binding to 14-3-3 proteins through phospho-Thr(649). We recently provided genetic evidence that AS160-Thr(649) phosphorylation/14-3-3 binding plays a key role in mediating insulin-stimulated glucose uptake in muscle. Contraction has also been proposed to increase phosphorylation of AS160 and TBC1D1 via AMPK, which could be detected by a generic phospho-Akt substrate (PAS) antibody. Here, analysis of AS160 immunoprecipitates from muscle extracts with site-specific phospho-antibodies revealed that contraction and AICAR caused no increase but rather a slight decrease in phosphorylation of the major PAS recognition site AS160-Thr(649). In line with this, contraction failed to enhance 14-3-3 binding to AS160. Consistent with previous reports, we also observed that in situ contraction stimulated the signal intensity of PAS antibody immunoreactive protein of similar to 150-160 kDa in muscle extracts. Using a TBC1D1 deletion mutant mouse, we showed that TBC1D1 protein accounted for the majority of the PAS antibody immunoreactive signals of similar to 150-160 kDa in extracts of contracted muscles. Consistent with the proposed role of AS160-Thr(649) phosphorylation/14-3-3 binding in mediating glucose uptake, AS160-Thr(649)Ala knock-in mice displayed normal glucose uptake upon contraction and AICAR in isolated muscles. We conclude that the previously reported PAS antibody immunoreactive band similar to 150-160 kDa, which were increased upon contraction, does not represent AS160 but TBC1D1, and that AS160-Thr(649)Ala substitution impairs insulin-but neither contraction- nor AICAR-stimulated glucose uptake in mouse skeletal muscle.</p>

AB - <p>Ducommun S, Wang HY, Sakamoto K, MacKintosh C, Chen S. Thr(649)Ala-AS160 knock-in mutation does not impair contraction/AICAR-induced glucose transport in mouse muscle. Am J Physiol Endocrinol Metab 302: E1036-E1043, 2012. First published February 7, 2012; doi:10.1152/ajpendo.00379.2011.-AS160 and its closely related protein TBC1D1 have emerged as key mediators for both insulin-and contraction-stimulated muscle glucose uptake through regulating GLUT4 trafficking. Insulin increases AS160 phosphorylation at multiple Akt/PKB consensus sites, including Thr(649), and promotes its binding to 14-3-3 proteins through phospho-Thr(649). We recently provided genetic evidence that AS160-Thr(649) phosphorylation/14-3-3 binding plays a key role in mediating insulin-stimulated glucose uptake in muscle. Contraction has also been proposed to increase phosphorylation of AS160 and TBC1D1 via AMPK, which could be detected by a generic phospho-Akt substrate (PAS) antibody. Here, analysis of AS160 immunoprecipitates from muscle extracts with site-specific phospho-antibodies revealed that contraction and AICAR caused no increase but rather a slight decrease in phosphorylation of the major PAS recognition site AS160-Thr(649). In line with this, contraction failed to enhance 14-3-3 binding to AS160. Consistent with previous reports, we also observed that in situ contraction stimulated the signal intensity of PAS antibody immunoreactive protein of similar to 150-160 kDa in muscle extracts. Using a TBC1D1 deletion mutant mouse, we showed that TBC1D1 protein accounted for the majority of the PAS antibody immunoreactive signals of similar to 150-160 kDa in extracts of contracted muscles. Consistent with the proposed role of AS160-Thr(649) phosphorylation/14-3-3 binding in mediating glucose uptake, AS160-Thr(649)Ala knock-in mice displayed normal glucose uptake upon contraction and AICAR in isolated muscles. We conclude that the previously reported PAS antibody immunoreactive band similar to 150-160 kDa, which were increased upon contraction, does not represent AS160 but TBC1D1, and that AS160-Thr(649)Ala substitution impairs insulin-but neither contraction- nor AICAR-stimulated glucose uptake in mouse skeletal muscle.</p>

U2 - 10.1152/ajpendo.00379.2011

DO - 10.1152/ajpendo.00379.2011

M1 - Article

JO - American Journal of Physiology, Endocrinology and Metabolism

JF - American Journal of Physiology, Endocrinology and Metabolism

SN - 0193-1849

IS - 9

VL - 302

SP - E1036-E1043

ER -

Documents

Library & Learning Centre

Contact | Accessibility | Policy