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Role and mechanism of homocysteine in affecting hepatic protein-tyrosine phosphatase 1B

Author:
Liu, Ping, Huang, Jin, Zhong, Liangwei
Source:
Biochimica et biophysica acta 2019 v.1863 no.5 pp. 941-949
ISSN:
0304-4165
Subject:
NADP (coenzyme), RNA interference, Western blotting, glucose, hepatocytes, homocysteine, insulin, insulin receptors, liquid chromatography, phosphorylation, precipitin tests, tandem mass spectrometry, thiols, thioredoxins
Abstract:
Elevated homocysteine is epidemiologically related to insulin resistance. Protein-tyrosine phosphatase 1B (PTP1B) is a negative regulator of insulin signaling. However, the effect of homocysteine on PTP1B remains unclear.S-homocysteinylated PTP1B was identified by LC-ESI-MS/MS. The ability of thioredoxin system to recover active PTP1B from S-homocysteinylated PTP1B was confirmed by RNA interference. To address the mechanism for homocysteine to affect PTP1B activity, we performed 5-IAF insertion, activity assays, Western blotting, co-immunoprecipitation and glucose uptake experiments.The thiol-containing form of homocysteine (HcySH) suppressed phosphorylation of insulin receptor-β subunit, but enhanced PTP1B activity. This phenomenon was partially related to the fact that HcySH promoted PTP1B expression. Although the disulfide-bonded form of homocysteine (HSSH) modified PTP1B to form an inactive S-homocysteinylated PTP1B, HcySH-induced increase in the activities of cellular thioredoxin and thioredoxin reductase, components of thioredoxin system, could recover active PTP1B from S-homocysteinylated PTP1B. Thioredoxin system transferred electrons from NADPH to S-homocysteinylated PTP1B, regenerating active PTP1B in vitro and in hepatocytes. The actions of HcySH were also related with decrease in hepatic glucose uptake.The effect of HcySH/HSSH on PTP1B activity depends, at least partially, on the ratio of active PTP1B and S-homocysteinylated PTP1B. High HcySH-induced an increase in thioredoxin system activity is beneficial to de-S-homocysteinylation and is good for PTP1B activity.Our data provide a novel insight into post-translational regulation of PTP1B, and expand the biological functions of thioredoxin system.
Agid:
6338548