Analysis Of Protein Ingredient In Urine Calcium Oxalate Stones

Objectives:To Explore the characteristics of Protein Ingredient in Urine Calcium O xalate Stones,for the subsequent further exploration within the protein’s role in stone formation and associated molecular mechanism.Methods:20 clinical stone were collected after surgery.Stones were analyzed by infrared spectroscopy to determine the components of stones.RIPA lysis buffer were used to dissolve stones,and protein sample was concentrated after freezing drying.Qproteome Bacterial Protein Prep K it were adopted to extract e.coli protein.Then we measured the protein concentration using Bradford method,and used gel electrophoresis to isolate and purify.The molecular weight of the protein was identified by using coomassie brilliant blue staining and silver staining,and compared the protein band patterns.The peptide fragment was extracted by gel enzyme solution technology,the protein components were identified by mass spectrom-etry.The results of mass spectrometry were analyzed by Gene Ontology.Results:1.The average protein concentration of calcium oxalate stone was 0.57ug/ uL.There was a relatively high protein content of calcium oxalate in urinary tract infection(p<0.05).E.coli protein concentration was 5.38ug/uL.2.SDS-PAGE electrophoresis results indicates that there are at least 3-4 protein bands in calcium oxalate stones,mainly concentrating in molecular 66.2-116.0KD,25.0-35.0 KD.The protein bands of e.coli are mainly concentrated in 35.0KDa-116.0KDa.The protein band pattern indicated that the protein composition of calcium oxalate stone and e.coli were overlapping.3.Protein Mass Spectrometry showed that there are 88 kinds of protein in the calcium oxalate stone,which were mainly binding proteins.Amo ng them,S100 protein family,fibronectin,keratin and etc,which may be associated with inflammation.onclusions:The protein composition of calcium oxalate stone is complex.It is not only intersected with e.coli protein,but also has inflammation-related proteins.It is indicated that the formation of calcium oxalate stone may be associated with urinary tract infection.Meanwhile,the role of inflammation protein in stone formation is still to be lucubrated.