Characterization of the human adipocyte proteome and reproducibility of protein abundance by one-dimensional gel electrophoresis and HPLC - ESI - MS/MS

Xitao Xie, Zhengping Yi, Benjamin Bowen, Cassandra Wolf, Charles R. Flynn, Sandeep Sinha, Lawrence J. Mandarino, Christian Meyer

    Research output: Contribution to journalArticle

    31 Citations (Scopus)

    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 ∼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.

    Original languageEnglish (US)
    Pages (from-to)4521-4534
    Number of pages14
    JournalJournal of Proteome Research
    Volume9
    Issue number9
    DOIs
    StatePublished - Sep 3 2010

    Fingerprint

    Proteome
    Electrophoresis
    Adipocytes
    Gels
    High Pressure Liquid Chromatography
    Proteins
    Fatty Acids
    Abdominal Subcutaneous Fat
    Adipokines
    Lipids
    Lipolysis
    Mitochondrial Proteins
    Mitochondria
    Glycolysis
    Oxidative stress
    Heat-Shock Proteins
    Tandem Mass Spectrometry
    Lipid Metabolism
    High performance liquid chromatography
    Type 2 Diabetes Mellitus

    Keywords

    • adipocyte
    • human
    • mitochondria
    • proteomics

    ASJC Scopus subject areas

    • Biochemistry
    • Chemistry(all)

    Cite this

    Characterization of the human adipocyte proteome and reproducibility of protein abundance by one-dimensional gel electrophoresis and HPLC - ESI - MS/MS. / Xie, Xitao; Yi, Zhengping; Bowen, Benjamin; Wolf, Cassandra; Flynn, Charles R.; Sinha, Sandeep; Mandarino, Lawrence J.; Meyer, Christian.

    In: Journal of Proteome Research, Vol. 9, No. 9, 03.09.2010, p. 4521-4534.

    Research output: Contribution to journalArticle

    Xie, Xitao ; Yi, Zhengping ; Bowen, Benjamin ; Wolf, Cassandra ; Flynn, Charles R. ; Sinha, Sandeep ; Mandarino, Lawrence J. ; Meyer, Christian. / Characterization of the human adipocyte proteome and reproducibility of protein abundance by one-dimensional gel electrophoresis and HPLC - ESI - MS/MS. In: Journal of Proteome Research. 2010 ; Vol. 9, No. 9. pp. 4521-4534.
    @article{32616860dd8944aa896f0b893e633312,
    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 ∼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.",
    keywords = "adipocyte, human, mitochondria, proteomics",
    author = "Xitao Xie and Zhengping Yi and Benjamin Bowen and Cassandra Wolf and Flynn, {Charles R.} and Sandeep Sinha and Mandarino, {Lawrence J.} and Christian Meyer",
    year = "2010",
    month = "9",
    day = "3",
    doi = "10.1021/pr100268f",
    language = "English (US)",
    volume = "9",
    pages = "4521--4534",
    journal = "Journal of Proteome Research",
    issn = "1535-3893",
    publisher = "American Chemical Society",
    number = "9",

    }

    TY - JOUR

    T1 - Characterization of the human adipocyte proteome and reproducibility of protein abundance by one-dimensional gel electrophoresis and HPLC - ESI - MS/MS

    AU - Xie, Xitao

    AU - Yi, Zhengping

    AU - Bowen, Benjamin

    AU - Wolf, Cassandra

    AU - Flynn, Charles R.

    AU - Sinha, Sandeep

    AU - Mandarino, Lawrence J.

    AU - Meyer, Christian

    PY - 2010/9/3

    Y1 - 2010/9/3

    N2 - 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 ∼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.

    AB - 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 ∼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.

    KW - adipocyte

    KW - human

    KW - mitochondria

    KW - proteomics

    UR - http://www.scopus.com/inward/record.url?scp=77956300476&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=77956300476&partnerID=8YFLogxK

    U2 - 10.1021/pr100268f

    DO - 10.1021/pr100268f

    M3 - Article

    C2 - 20812759

    AN - SCOPUS:77956300476

    VL - 9

    SP - 4521

    EP - 4534

    JO - Journal of Proteome Research

    JF - Journal of Proteome Research

    SN - 1535-3893

    IS - 9

    ER -