Proteome Identification And Analysis Of Hepatocyte Growth-Promoting Factors By Mass Spectrometry

Hepatocyte Growth-Promoting Factors(PHGF)is the first one of first class national new biochemical drug in our country.It is a mixture of proteins and peptides,which are extracted and purified from fresh liver of anthony pig.PHGF is used in therapy of hepatic disease such as hepatitis and liver cirrhosis.So far,PHGF still plays an important role on hepatic disease treatment,with further improved preparation method and reproducibility control.However,the detail protein/peptide sequence and abundance,the active components in PHGF are still unrevealed and not reported yet,which lead to the lack of pharmacokinetics test and quality control of PHGF.In this study,we used the developed proteomics method based on LC-MS/MS technology to identify the protein/peptides sequence and abundance in PHGF thoroughly for first time,and analyze the protein function and pathway in order to reveal the correlation between actual sequences and related function/pathway of the proteins.The proteomics study of PHGF can provide important dataon the pharmacological efficacy information,and further improve the possible therapeutic mechanism of hepatic disease.The proteomics data is also helpful for the further research and production improvement of PHGF,as a good reference and direction.A total of 152 proteins/peptides were identified in PHGF by proteomics analysis based on LC-MS/MS.All of these proteins have never been reported in PHGF study,including some relative high abundant proteins,such as Hemoglobin,Malate dehydrogenase,Catalase,Fructose-1,6-bisphosphatase,Betaine-homocysteine S-methyltransferase,liver carboxylesterase,Aldehyde dehydrogenase,Epoxide hydrolase and ATP synthase subunit alpha.The properties of these proteins were likely relevant to the activity of promoting hepatocyte growth.Other relative low abundant proteins were also important for potential regulation in the activity of PHGF,coordinating the therapeutic effect.Many key molecular functions were annotated and clustered from the identified proteins,including electron carrier activity,protein binding,structural constituent of ribosome,heme binding,flavin adenine dinucleotide binding,fatty-acyl-CoA binding,succinate dehydrogenase activity,poly(A)RNA binding and NADP binding activity.Pathway analysis on KEGG of the proteins also showed relative pathways,such as Biosynthesis of antibiotics,Carbon metabolism,Metabolic pathways,Citrate cycle,Biosynthesis of amino acids,Propanoate metabolism,Cysteine and methionine metabolism,Glycolysis / Gluconeogenesis,Valine,leucine and isoleucine degradation and Fatty acid degradation.All these clustered enriched molecular functions and pathways have potential to improve the activity of promoting hepatocyte growth,providing important database for PHGF regulation research.The result of this study makes good contribution to further understand the functions of proteins/peptides,production improvement,pharmacokinetics research,drug quality control and medication optimization of PHGF as the important reference dataset.On the other hand,this study also shows that the advanced proteomics method is powerful for protein/peptide drug research,and is very useful for therapy of more diseases,drug development and drug production.