Nakashima Yoshiki, Miyagi-Shiohira Chika, Kobayashi Naoya, Saitoh Issei, Watanabe Masami, Noguchi Hirofumi
a Department of Regenerative Medicine , Graduate School of Medicine, University of the Ryukyus , Okinawa , Japan.
b Okayama Saidaiji Hospital , Okayama , Japan.
Islets. 2017 Nov 2;9(6):159-176. doi: 10.1080/19382014.2017.1389826. Epub 2017 Nov 3.
Liquid chromatography with tandem mass spectrometry (LC-MS/MS) is a proteome analysis method, and the shotgun analysis by LC-MS/MS comprehensively identifies proteins from tissues and cells with high resolving power. In this study, we analyzed the protein expression in pancreatic tissue by LC-MS/MS. Islets isolated from porcine pancreata (purity ≥95%) and exocrine tissue (purity ≥99%) were used in this study. LC-MS/MS showed that 13 proteins were expressed in pancreatic islets only (Group I), 43 proteins were expressed in both islets and exocrine tissue (Group I&E), and 102 proteins were expressed in exocrine tissue only (Group E). Proteins involved in islet differentiation and cell proliferation were identified in Group I (e.g. CLUS, CMGA, MIF). In addition, various functional proteins (e.g. SCG2, TBA1A) were identified in islet by using the new method of 'principal component analysis (PCA)'. However, the function of such proteins on islets remains unclear. EPCAM was identified in Group E. Group E was found to include proteins involved in clinical inflammatory diseases such as pancreatitis (e.g. CBPA1, CGL, CYTB, ISK1 and PA21B). Many of these identified proteins were reported less frequently in previous studies, and HS71B, NEC2, PRAF3 and SCG1 were newly detected in Group I while CPNS1, DPEP1, GANAB, GDIB, GGT1, HSPB1, ICTL, VILI, MUTA, NDKB, PTGR1, UCHL3, VAPB and VINC were newly detected in Group E. These results show that comprehensive expression analysis of proteins by LC-MS/MS is useful as a method to investigate new factors constructing cellular component, biological process, and molecular function.
液相色谱-串联质谱法（LC-MS/MS）是一种蛋白质组分析方法，通过LC-MS/MS进行的鸟枪法分析能够以高分辨率全面鉴定组织和细胞中的蛋白质。在本研究中，我们通过LC-MS/MS分析了胰腺组织中的蛋白质表达情况。本研究使用了从猪胰腺分离出的胰岛（纯度≥95%）和外分泌组织（纯度≥99%）。LC-MS/MS结果显示，仅在胰岛中表达的蛋白质有13种（第一组），在胰岛和外分泌组织中均表达的蛋白质有43种（第一组和外分泌组织组），仅在外分泌组织中表达的蛋白质有102种（外分泌组织组）。在第一组中鉴定出了参与胰岛分化和细胞增殖的蛋白质（如CLUS、CMGA、MIF）。此外，通过“主成分分析（PCA）”新方法在胰岛中鉴定出了各种功能蛋白质（如SCG2、TBA1A）。然而，这些蛋白质在胰岛上的功能仍不清楚。在外分泌组织组中鉴定出了EPCAM。外分泌组织组被发现包含参与胰腺炎等临床炎症性疾病的蛋白质（如CBPA1、CGL、CYTB、ISK1和PA21B）。这些鉴定出的蛋白质中有许多在先前的研究中报道较少，在第一组中新检测到了HS71B、NEC2、PRAF3和SCG1，而在外分泌组织组中新检测到了CPNS1、DPEP1、GANAB、GDIB、GGT1、HSPB1、ICTL、VILI、MUTA、NDKB、PTGR1、UCHL3、VAPB和VINC。这些结果表明，通过LC-MS/MS对蛋白质进行全面表达分析作为一种研究构建细胞成分、生物学过程和分子功能的新因素的方法是有用的。
