Research resource: identification of novel growth hormone-regulated phosphorylation sites by quantitative phosphoproteomics.

Molecular Endocrinology
Bridgette N RayChristin Carter-Su

Abstract

GH and GH receptors are expressed throughout life, and GH elicits a diverse range of responses, including growth and altered metabolism. It is therefore important to understand the full spectrum of GH signaling pathways and cellular responses. We applied mass spectrometry-based phosphoproteomics combined with stable isotope labeling with amino acids in cell culture to identify proteins rapidly phosphorylated in response to GH in 3T3-F442A preadipocytes. We identified 132 phosphosites in 95 proteins that exhibited rapid (5 or 15 min) GH-dependent statistically significant increases in phosphorylation by more than or equal to 50% and 96 phosphosites in 46 proteins that were down-regulated by GH by more than or equal to 30%. Several of the GH-stimulated phosphorylation sites were known (e.g. regulatory Thr/Tyr in Erks 1 and 2, Tyr in signal transducers and activators of transcription (Stat) 5a and 5b, Ser939 in tuberous sclerosis protein (TSC) 2 or tuberin). The remaining 126 GH-stimulated sites were not previously associated with GH. Kyoto Encyclopedia of Genes and Genomes pathway analysis of GH-stimulated sites indicated enrichment in proteins associated with the insulin and mammalian target of rapamycin (mTOR) pathways, regulat...Continue Reading

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Citations

Jan 11, 2014·BMC Cell Biology·Dhanshri KakadeOlasunkanmi A J Adegoke
Oct 25, 2016·Clinical Medicine Insights. Endocrinology and Diabetes·Jesús DevesaPablo Devesa
May 14, 2016·Molecular Endocrinology·Elena Pedraz-CuestaStine F Pedersen
Feb 8, 2019·JCI Insight·Vera ChesnokovaShlomo Melmed
Nov 2, 2019·Prostaglandins & Other Lipid Mediators·Marco RahmStefanie M Hauck
Aug 28, 2014·Journal of Proteome Research·Michael ReinartzAxel Gödecke

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Methods Mentioned

BETA
protein assay
nuclear translocation
Immunoprecipitation

Software Mentioned

mouse
Mascot
LI
Odyssey
COR
MaxQuant

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