Identification And Analysis Of Mass Spectra For Calcium Oxalate And Uric Acid Kidney Stones Matrix Proteins

Objectives:To Explore the different composition of proteins in the urinary kidney stones, for the subsequent further exploration within the protein’s role in stone formation and associated molecular mechanism. Methods:Firstly,we collected 150 clinical stone between June 2013 and December 2014 in our hospital about 0.5cm-4.8cm in large after surgery of percutaneous nephrolithotomy, ureteroscopy and laparoscopic, etc. After washing, drying, grinding,stones was analyzed by infrared spectroscopy. While using RIPA lysis buffer to dissolve stones, protein sample was concentrated after freezing drying, then the protein samples were isolated and purified, by gel electrophoresis The obtained proteins were analyzed with reverse-phase liquid chromatography–tandem MS. The acquired data were searched against a Swiss Prot human protein database using Matrix Science, Mascot. The identified proteins were submitted to the Ami GO Web site for gene ontology analysis. They were also sumitted to Metacore software and Kyoto Encyclopedia of Genes and Genomes website(KEGG) for pathway analysis. Results:1. In the 150 cases of urinary tract stones, there are 33 pure calcium oxalate stones, 12 uric acid stones, 66 cases of calcium oxalate stones with Calcium Carbonate, 19 cases of calcium oxalate combined with uric acid stones and 20 cases of calcium oxalate combined with Calcium phosphate2.SDS-PAGE electrophoresis results table: In 33 cases of calcium oxalate stones there are 3-4 protein bands, mainly in molecular 70-100 KD, 17-26 KD area; while 12 cases of uric acid stones within 5-6 protein bands, mainly in 70-100 KD, 30-40 KD, 17-26 KD area.3. Protein MS Results: There are 88 kinds of protein in the calcium oxalate stone, and 283 kinds of protein in the uric acid stones. GO analysis, pathway results show that: binding proteins in the stones, accounting for 45.8% of the total protein in the calcium oxalate stone, while 44.34% in the uric acid stones. Among them, S100A8, S100A9, fibronectin, keratin and other inflammatory protein may be related to calculi forming. Conclusions:1. protein composition and content vary in different kind of stones.2. protein composition and content in the same kind of stones vary slightly3. There are 88 kinds of protein in the calcium oxalate stone, 283 kinds of protein in the uric acid stone.