title: Profiling and semiquantitative analysis of the cell surface proteome in human mesenchymal stem cells.
abstract: Mulitpotent mesenchymal stem cells (MSCs) derived from human bone marrow are promising candidates for the development of cell therapeutic strategies. MSC surface protein profiles provide novel biological knowledge concerning the proliferation and differentiation of these cells, including the potential for identifying therapeutic targets. Basic fibroblast growth factor (bFGF) affects cell surface proteins, which are associated with increased growth rate, differentiation potential, as well as morphological changes of MSCs in vitro. Cell surface proteins were isolated using a biotinylation-mediated method and identified using a combination of one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and mass spectrometry. The resulting gel lines were cut into 20 bands and digested with trypsin. Each tryptic fragment was analyzed by liquid chromatography-electrospray ionization tandem mass spectrometry. Proteins were identified using the Mascot search program and the International Protein Index human database. Noble MSC surface proteins (n = 1,001) were identified from cells cultured either with (n = 857) or without (n = 667) bFGF-containing medium in three independent experiments. The proteins were classified using FatiGO to elucidate their function. We also confirmed the proteomics results using Western blotting and immunofluorescence microscopic analysis. The nature of the proteins identified makes it clear that MSCs express a wide variety of signaling molecules, including those related to cell differentiation. Among the latter proteins, four Ras-related Rab proteins, laminin-R, and three 14-3-3 proteins that were fractionated from MSCs cultured on bFGF-containing medium are implicated in bFGF-induced signal transduction of MSCs. Consequently, these finding provide insight into the understanding of the surface proteome of human MSCs.
authors: Mass Spectrometry Research Center, Korea Basic Science Institute, Ochang 363-887, Republic of Korea.
authors: Sang Kwang
authors: SK
authors: Lee
authors: Jae Ho
authors: JH
authors: Kim
authors: Sung-Soo
authors: SS
authors: Kim
authors: Taewook
authors: T
authors: Kang
authors: Nam Hyun
authors: NH
authors: Park
authors: Kyung-Hoon
authors: KH
authors: Kwon
authors: Sang Sook
authors: SS
authors: Lee
authors: Zee Won
authors: ZW
authors: Lee
authors: Hae young
authors: Hy
authors: Suh-Kim
authors: Kun
authors: K
authors: Cho
authors: Su Yeoung
authors: SY
authors: Yun
authors: Ji Young
authors: JY
authors: Han
authors: Jong Shin
authors: JS
authors: Yoo
authors: Hyun Joo
authors: HJ
authors: An
authors: Young Mok
authors: YM
authors: Park
mesh_terms: D048948:14-3-3 Proteins / Q000032:analysis / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D039902:Multipotent Stem Cells / Q000378:metabolism; D011506:Proteins / Q000032:analysis* / Q000145:classification / Q000378:metabolism; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D017474:Receptors, Laminin / Q000378:metabolism; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry
chemical_list: D048948:14-3-3 Proteins; D011506:Proteins; D020543:Proteome; D017474:Receptors, Laminin; D016222:Fibroblast Growth Factor 2