Membrane proteomic analysis of human mesenchymal stromal cells during adipogenesis.
Mesenchymal stromal cells (MSCs) have proven useful for cell and immune therapy, but the molecular constituents responsible for their functionalities, in particular, those on the plasma membrane, remain largely unknown. Here we employed both gel and nongel based MS to analyze human MSCs' membrane proteome before and after adipogenesis. 2-DE of cells that were pretreated with membrane impermeable fluorescent dyes revealed that both the whole cell proteome and the cell surface subproteome were independent of donors. LC coupled with tandem MS analysis of the plasma membrane-containing fraction allowed us to identify 707 proteins, approximately half of which could be annotated as membrane-related proteins. Of particular interest was a subset of ectodomain-containing membrane-bound proteins that encompass most known surface markers for MSCs, but also contain a multitude of solute carriers and ATPases. Upon adipogenic differentiation, this proteomic profile was amended to include several proteins involved in lipid metabolism and trafficking, at the expense of, most noticeably, ectoenzymes. Our results here provide not only a basis for future studies of MSC-specific molecular mechanisms, but also a molecular inventory for the development of antibody-based cell isolation and identification procedures.
Jeong
Ju Ah
JA
Biotherapeutic Division, GenExel-Sein Inc., Daejon, Korea.
Ko
Kyung-Min
KM
Park
Hyung Soon
HS
Lee
Jinsook
J
Jang
Cholsoon
C
Jeon
Choon-Ju
CJ
Koh
Gou Young
GY
Kim
Hoeon
H
D050156:Adipogenesis / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D002478:Cells, Cultured; D002853:Chromatography, Liquid / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D020869:Gene Expression Profiling; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D040901:Proteomics; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods
D008565:Membrane Proteins; D012333:RNA, Messenger