NDLI: Identification of cytoplasmic and membrane-associated complexes in human embryonic stem cells using blue native PAGE

Content Provider	Royal Society of Chemistry (RSC)
Author	Pan, Tai-Long Shekari, Faezeh Wang, Pei-Wen Salekdeh, Ghasem Hosseini Baharvand, Hossein Taei, Adele
Copyright Year	2011
Abstract	Human embryonic stem cells (hESCs) have great potential for use in developmental biology studies, functional genomics applications, drug screening, and regenerative medicine. A detailed understanding of the molecular mechanisms that are responsible for maintaining the undifferentiated and pluripotent nature of hESCs is essential for their effective therapeutic application. It has become evident that many complex cellular processes are carried out by assemblies of protein molecules (protein complexes). Blue native polyacrylamide gel electrophoresis (BN-PAGE) has been used to separate protein complexes from whole cell lysates. Using BN-PAGE, we resolved cytoplasmic and membrane-associated complexes from hESCs and characterised their composition, stoichiometry, and dynamics by denaturing SDS-PAGE. The reliability of the fractionation was examined by western blot analysis of membrane and cytosolic markers. MALDI TOF/TOF mass spectrometry identified 119 cytosolic and 69 membrane proteins from the BN-PAGE proteome maps. Potential protein complexes were validated by computational prediction of possible protein–protein interactions using the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) database. Based on BN-PAGE gels and validation by databases, 82 heteromultimeric and 47 homomultimeric protein complexes have been found in hESCs. Resolving some of the protein complexes provided insight into the function of previously uncharacterised complexes in hESCs.
Starting Page	2688
Ending Page	2701
Page Count	14
File Format	HTM / HTML PDF
ISSN	1742206X
Volume Number	7
Issue Number	9
Journal	Molecular BioSystems
DOI	10.1039/c1mb05135k
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	Western blot Polyacrylamide gel electrophoresis Stoichiometry Regenerative medicine Gel electrophoresis Ionization Cell potency Proteome Molecular biology Mass spectrometry Functional genomics Protein
Content Type	Text
Resource Type	Article
Subject	Molecular Biology Biotechnology

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