Differential Proteomics Analysis Of High Quality Brush Hair In Yangtze River Delta White Goat

Yangtze River Delta white goat is a goat breed which can produce high quality brush hair (namely Type Ⅲ Hair) around the world. The Type Ⅲ Hair is used for making high-grade writing brush for its white color, straight hair shaft, long and fine tip, bright luster and good flexibility. Because of its unique style, the writing brush made of Type Ⅲ hair wins popular praise, together with ink stick, ink slab and rice paper to be known as "the scholar’s four jewels". As the source of high-grade writing brush, the Type Ⅲ Hair has precious value.In this study, the emphasis was on the research of the formation mechanisms of high quality brush hair at protein level, constructing Type Ⅲ Hair and non-Type Ⅲ Hair goat skin two-dimensional gel electrophoresis (2-DE) maps, screening and identifying differential expressed proteins. The aim was searching for Type Ⅲ Hair-associated proteins, and forecasting gene sequences through amino acid sequences, which could lay the foundation of Type Ⅲ Hair-associated gene clone and the further molecular breeding.We collected six to eight months old Type Ⅲ Hair and non-Type Ⅲ Hair goats’ skin tissues. Total proteins were extracted,1000μg protein was utilized for isoelectric focusing (IEF), after immobilized pH gradient (IPG) strip equilibration, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was carried out. The gel was visualized by coomassie brilliant blue stain. After scanned, the differential expressed proteins were found by PDquest8.0software. Peptide MS and MS/MS were performed on an Ultraflex Ⅲ TOF/TOF mass spectrometer, the MS and MS/MS data were integrated by Mascot2.3(Matrix Science) for protein identification.There were some results as follows.(1) Type Ⅲ Hair and non-Type Ⅲ Hair goat skin2-DE maps were constructed, respectively.(2)33differential expressed protein spots were found by PDquest8.0software, in which20differential expressed protein spots were up regulated in Type Ⅲ Hair goat skin, and the other13differential expressed protein spots were up regulated in non-Type Ⅲ Hair goat skin.(3) Ten peptide mass fingerprintings (PMF) were obtained by Ultraflex Ⅲ TOF/TOF mass spectrometer.(4) Through matching based on Ultraflex III TOF/TOF and MASCOT search engine, four differential expressed proteins, Fibrinogen Beta Chain Isoform1, ATP Synthase Beta Subunit, Succinyl-CoA:3-Ketoacid-Coenzyme A Transferase1-Mitochondrial-Like and Actin-Cytoplasmic1were identified. The two proteins in front were up regulated in Type Ⅲ Hair goat skin, and the latter two proteins were up regulated in non-Type Ⅲ Hair goat skin.The analysis results were as follows.(1) Fibrinogen Beta Chain Isoform1could offer nutrient for hair follicle and participate in cortical cell adhesion, and proliferation to promote the formation of cortex.(2) ATP Synthase Beta Subunit could offer energy for the synthesis of keratin cells through accelerating the synthesis and hydrolysis of ATP. The increasement of cortex contents was possible when the energy was too much, which could promote the formation of no-medullary segment.(3) Succinyl-CoA:3-Ketoacid-Coenzyme A Transferase1-Mitochondrial-Like could affect the formation of hair shaft through androgen and promote the synthesis of highly expressed proteins in medulla to facilitate the formation of medulla.(4) Actin-Cytoplasmic1interacted with keratins to improve the morphology of hair follicle associated cells, it played an important role in maintaining the morphology of corneous trabeculae and restricting the movement of keratinocyte’membrane proteins in medulla, which would be useful to the formation of vesicular structural medulla.This study constructed high-resolution and good repeatability Type Ⅲ Hair and non-Type Ⅲ Hair goat skin2-DE maps, identified some Type Ⅲ Hair-associated differential expressed proteins, which laid a good foundation for further study on the formation mechanisms of high quality brush hair in Yangtze River Delta white goat.