title: Characterization of the Human Adipocyte Proteome and Reproducibility of Protein Abundance by One-Dimensional Gel Electrophoresis and HPLC-ESI-MS/MS.
abstract: Abnormalities in adipocytes play an important role in various conditions, including the metabolic syndrome, type 2 diabetes mellitus and cardiovascular disease, but little is known about alterations at the protein level. We therefore sought to (1) comprehensively characterize the human adipocyte proteome for the first time and (2) demonstrate feasibility of measuring adipocyte protein abundances by one-dimensional SDS-PAGE and high performance liquid chromatography-electron spray ionization-tandem mass spectrometry (HPLC-ESI-MS/MS). In adipocytes isolated from approximately 0.5 g of subcutaneous abdominal adipose tissue of three healthy, lean subjects, we identified a total of 1493 proteins. Triplicate analysis indicated a 22.5% coefficient of variation of protein abundances. Proteins ranged from 5.8 to 629 kDa and included a large number of proteins involved in lipid metabolism, such as fatty acid transport, fatty acid oxidation, lipid storage, lipolysis, and lipid droplet maintenance. Furthermore, we found most glycolysis enzymes and numerous proteins associated with oxidative stress, protein synthesis and degradation as well as some adipokines. 22% of all proteins were of mitochondrial origin. These results provide the first detailed characterization of the human adipocyte proteome, suggest an important role of adipocyte mitochondria, and demonstrate feasibility of this approach to examine alterations of adipocyte protein abundances in human diseases.
authors.affiliation: Center for Metabolic Biology, Arizona State University, Tempe, Arizona, USA.
authors.forename: Xitao
authors.initials: X
authors.lastname: Xie
authors.forename: Zhengping
authors.initials: Z
authors.lastname: Yi
authors.forename: Benjamin
authors.initials: B
authors.lastname: Bowen
authors.forename: Cassandra
authors.initials: C
authors.lastname: Wolf
authors.forename: Charles R
authors.initials: CR
authors.lastname: Flynn
authors.forename: Sandeep
authors.initials: S
authors.lastname: Sinha
authors.forename: Lawrence J
authors.initials: LJ
authors.lastname: Mandarino
authors.forename: Christian
authors.initials: C
authors.lastname: Meyer
mesh_terms: D050153:Abdominal Fat / Q000166:cytology; D052438:Adipocytes, White / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006801:Humans; D053858:Metabolic Networks and Pathways; D051379:Mice; D011506:Proteins / Q000737:chemistry / Q000378:metabolism; D020543:Proteome / Q000737:chemistry*; D040901:Proteomics / Q000379:methods; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry
chemical_list: D011506:Proteins; D020543:Proteome
references.citation: J Biol Chem. 2004 Nov 5;279(45):46835-42
references.pmid: 15337753
references.citation: Mol Cell Proteomics. 2007 Jul;6(7):1257-73
references.pmid: 17409382
references.citation: J Leukoc Biol. 2008 Apr;83(4):804-16
references.pmid: 18252866
references.citation: J Nutr. 2004 Apr;134(4):762-70
references.pmid: 15051823
references.citation: Nat Cell Biol. 2007 Nov;9(11):1286-93
references.pmid: 17922004
references.citation: Proteomics. 2009 Feb;9(4):914-21
references.pmid: 19160396
references.citation: J Appl Physiol (1985). 1998 Feb;84(2):479-85
references.pmid: 9475856
references.citation: Expert Rev Proteomics. 2008 Dec;5(6):827-39
references.pmid: 19086862
references.citation: J Proteome Res. 2009 Apr;8(4):1682-93
references.pmid: 19714809
references.citation: Cell Signal. 2006 Apr;18(4):401-8
references.pmid: 16182514
references.citation: J Biol Chem. 2001 Sep 7;276(36):34182-8
references.pmid: 11447218
references.citation: Am J Clin Nutr. 2009 Jan;89(1):407-15
references.pmid: 19056584
references.citation: J Proteome Res. 2007 Aug;6(8):3256-65
references.pmid: 17608402
references.citation: Mol Cell Biol. 2003 Feb;23(3):1085-94
references.pmid: 12529412
references.citation: Mol Cell Proteomics. 2005 Apr;4(4):492-522
references.pmid: 15695426
references.citation: Nature. 2008 May 29;453(7195):657-61
references.pmid: 18408709
references.citation: Diabetes. 2008 Sep;57(9):2438-44
references.pmid: 18567819
references.citation: Bioinformatics. 2004 Jul 22;20(11):1785-94
references.pmid: 15037509
references.citation: Diabetes Metab. 2004 Sep;30(4):294-309
references.pmid: 15525872
references.citation: Mol Biosyst. 2007 May;3(5):354-60
references.pmid: 17460794
references.citation: J Clin Invest. 2004 Nov;114(9):1281-9
references.pmid: 15520860
references.citation: Proc Natl Acad Sci U S A. 1997 Jan 21;94(2):514-9
references.pmid: 9012815
references.citation: Eur J Endocrinol. 2003 Mar;148(3):293-300
references.pmid: 12611609
references.citation: Proteomics. 2003 Oct;3(10):1912-9
references.pmid: 14625853
references.citation: Am J Pathol. 2009 Sep;175(3):927-39
references.pmid: 19700756
references.citation: Mol Cell Endocrinol. 2008 Sep 10;291(1-2):63-70
references.pmid: 18565643
references.citation: Curr Biol. 2001 Jun 5;11(11):R446-9
references.pmid: 11516669
references.citation: J Proteome Res. 2005 Mar-Apr;4(2):570-7
references.pmid: 15822936
references.citation: J Clin Invest. 2002 Dec;110(12):1773-82
references.pmid: 12488427
references.citation: FEBS Lett. 2009 Dec 17;583(24):3966-73
references.pmid: 19850042
references.citation: Diabetes. 2005 May;54(5):1392-9
references.pmid: 15855325
references.citation: Diabetes. 2010 Jan;59(1):33-42
references.pmid: 19833877
references.citation: J Cell Biol. 2004 Jun 21;165(6):813-22
references.pmid: 15197175
references.citation: Nature. 2005 Jul 21;436(7049):356-62
references.pmid: 16034410
references.citation: J Lipid Res. 2007 Dec;48(12):2547-59
references.pmid: 17878492
references.citation: J Proteome Res. 2009 Dec;8(12):5532-40
references.pmid: 19827857
references.citation: J Lipid Res. 1994 Feb;35(2):177-93
references.pmid: 8169522
references.citation: FEBS Lett. 1981 Sep 28;132(2):235-8
references.pmid: 7028513
references.citation: Proteomics. 2003 Jan;3(1):36-44
references.pmid: 12548632
references.citation: Proteomics Clin Appl. 2007 Oct;1(10):1306-15
references.pmid: 21136627
references.citation: Biochem J. 1983 Jul 15;214(1):203-7
references.pmid: 6351848
references.citation: Annu Rev Nutr. 2007;27:79-101
references.pmid: 17313320
references.citation: Hum Reprod. 2008 Mar;23(3):651-61
references.pmid: 18156650
references.citation: Gene. 2004 Sep 29;340(1):141-50
references.pmid: 15556302
references.citation: Acta Physiol Scand. 2005 Aug;184(4):285-93
references.pmid: 16026420
references.citation: Mol Cell Proteomics. 2007 Jan;6(1):18-28
references.pmid: 17018519
references.citation: J Biol Chem. 2004 Dec 31;279(53):55137-46
references.pmid: 15496407
references.citation: Int J Obes Relat Metab Disord. 2003 Aug;27(8):875-88
references.pmid: 12861227
references.citation: FEBS Lett. 2006 Oct 9;580(23):5484-91
references.pmid: 16962104
references.citation: Nat Rev Mol Cell Biol. 2006 May;7(5):373-8
references.pmid: 16550215
references.citation: Cell. 2004 Jan 23;116(2):337-50
references.pmid: 14744442
references.citation: FASEB J. 2004 Jan;18(1):215-7
references.pmid: 14630696
references.citation: Diabetes. 1996 Nov;45(11):1644-54
references.pmid: 8866574
references.citation: Diabetes. 2007 Jul;56(7):1751-60
references.pmid: 17456854
references.citation: Electrophoresis. 2000 Oct;21(16):3345-50
references.pmid: 11079554
references.citation: Proteomics. 2004 Feb;4(2):438-41
references.pmid: 14760714
references.citation: Mol Cell Proteomics. 2008 Apr;7(4):631-44
references.pmid: 18029349
references.citation: J Cell Sci. 2009 Mar 15;122(Pt 6):749-52
references.pmid: 19261844
references.citation: Nat Struct Mol Biol. 2004 Nov;11(11):1044-8
references.pmid: 15523480
references.citation: Trends Immunol. 2009 May;30(5):201-8
references.pmid: 19356981
references.citation: J Biol Chem. 2005 May 13;280(19):19146-55
references.pmid: 15731108
references.citation: J Appl Physiol (1985). 2007 Apr;102(4):1671-6
references.pmid: 17082366
references.citation: Mol Cell Proteomics. 2007 Apr;6(4):589-600
references.pmid: 17255083
references.citation: Endocrinology. 2008 Mar;149(3):942-9
references.pmid: 18202123
references.citation: Ann N Y Acad Sci. 2004 Dec;1030:508-36
references.pmid: 15659836