Proteomics

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Quantitative proteomics and phosphoproteomics analysis revealed different regulatory mechanisms of Halothane and Rendement Napole genes in porcine muscle metabolism


ABSTRACT: Pigs with the Halothane (HAL) and Rendement Napole (RN) gene mutations have different muscle energy metabolism patterns. It is not completely understood how HAL and RN mutations regulate glucose and energy metabolism in muscle. In order to investigate potential signaling pathways and phosphorylation events regulating energy metabolism postmortem, muscle samples were collected at 45 min exsanguination from four genotypes of pigs: wildtype, RN, HAL and RN-HAL double mutations and subjected to quantitative proteomic and phosphoproteomic analysis. The study led to the identification of 932 proteins from the non-modified peptide fractions and 1885 phosphoproteins with 9619 phosphorylation sites from the enriched phosphopeptide fractions. Among them, 128 proteins at total protein level and 323 phosphosites from 91 phosphoproteins were significantly regulated in at least one genotype compared to other genotypes. The quantitative analysis revealed that the RN mutation mainly affected the protein expression abundance in muscle. Specifically, it greatly increased the abundance of proteins related to mitochondrial respiratory chain and energy metabolism, thereby enhancing the muscle oxidative capacity. The higher content of UGP2 in RN mutant animals may contribute to high glycogen storage. However, presence of the HAL mutation resulted in major effects on the regulation of protein phosphorylation. Most affected phosphoproteins in HAL mutant related muscle were determined with up-regulated phosphorylation sites, and related to calcium signaling, muscle contraction, glycogen, glucose and energy metabolism, and cellular stress. The increase phosphorylation of CAMK2 was detected in HAL mutation, it may lead to activation of CAMK2 as well as its downstream signaling cascades. The phosphorylation changes may contribute to accelerated glyocogen metabolism, acute stress response, muscle fiber hypertrophy and hyperacute rigor mortis in HAL mutant animals. Our findings identify a set of protein targets in two disparate pig mutations known to alter muscle energy metabolism and provide additional insight into the regulatory mechanisms underlying postmortem energy metabolism in pig muscle.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Sus Scrofa Domesticus (domestic Pig)

TISSUE(S): Skeletal Muscle Fiber

SUBMITTER: Honggang Huang  

LAB HEAD: Rene Lametsch

PROVIDER: PXD008292 | Pride | 2018-06-21

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
FTARLA56-68.msf Msf
FTBRLA69-81.msf Msf
FTCRLA82-94.msf Msf
PhosphoA-RLA15-28.msf Msf
PhosphoB-RLA29-42.msf Msf
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Publications

Quantitative Proteomics and Phosphoproteomics Analysis Revealed Different Regulatory Mechanisms of Halothane and Rendement Napole Genes in Porcine Muscle Metabolism.

Huang Honggang H   Scheffler Tracy L TL   Gerrard David E DE   Larsen Martin R MR   Lametsch René R  

Journal of proteome research 20180629 8


Pigs with the Halothane (HAL) or Rendement Napole (RN) gene mutations demonstrate abnormal muscle energy metabolism patterns and produce meat with poor quality, classified as pale, soft, and exudative (PSE) meat, but it is not well understood how HAL and RN mutations regulate glucose and energy metabolism in porcine muscle. To investigate the potential signaling pathways and phosphorylation events related to these mutations, muscle samples were collected from four genotypes of pigs, wild type, R  ...[more]

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