A proteomic approach for identification of secreted proteins during the differentiation of 3T3-L1 preadipocytes to adipocytes

Irina Kratchmarova; Dario E Kalume; Blagoy Blagoev; Philipp E Scherer; Alexandre V Podtelejnikov; Henrik Molina; Perry E Bickel; Jens S Andersen; Minerva M Fernandez; Jacob Bunkenborg; Peter Roepstorff; Karsten Kristiansen; Harvey F Lodish; Matthias Mann; Akhilesh Pandey

A proteomic approach for identification of secreted proteins during the differentiation of 3T3-L1 preadipocytes to adipocytes

Irina Kratchmarova, Dario E Kalume, Blagoy Blagoev, Philipp E Scherer, Alexandre V Podtelejnikov, Henrik Molina, Perry E Bickel, Jens S Andersen, Minerva M Fernandez, Jacob Bunkenborg, Peter Roepstorff, Karsten Kristiansen, Harvey F Lodish, Matthias Mann, Akhilesh Pandey

227 Citations (Scopus)

Abstract

We have undertaken a systematic proteomic approach to purify and identify secreted factors that are differentially expressed in preadipocytes versus adipocytes. Using one-dimensional gel electrophoresis combined with nanoelectrospray tandem mass spectrometry, proteins that were specifically secreted by 3T3-L1 preadipocytes or adipocytes were identified. In addition to a number of previously reported molecules that are up- or down-regulated during this differentiation process (adipsin, adipocyte complement-related protein 30 kDa, complement C3, and fibronectin), we identified four secreted molecules that have not been shown previously to be expressed differentially during the process of adipogenesis. Pigment epithelium-derived factor, a soluble molecule with potent antiangiogenic properties, was found to be highly secreted by preadipocytes but not adipocytes. Conversely, we found hippocampal cholinergic neurostimulating peptide, neutrophil gelatinase-associated lipocalin, and haptoglobin to be expressed highly by mature adipocytes. We also used liquid chromatography-based separation followed by automated tandem mass spectrometry to identify proteins secreted by mature adipocytes. Several additional secreted proteins including resistin, secreted acidic cysteine-rich glycoprotein/osteonectin, stromal cell-derived factor-1, cystatin C, gelsolin, and matrix metalloprotease-2 were identified by this method. To our knowledge, this is the first study to identify several novel secreted proteins by adipocytes by a proteomic approach using mass spectrometry.

Original language	English
Journal	Molecular and Cellular Proteomics
Volume	1
Issue number	3
Pages (from-to)	213-22
Number of pages	9
ISSN	1535-9476
Publication status	Published - 2002
Externally published	Yes

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Kratchmarova, I., Kalume, D. E., Blagoev, B., Scherer, P. E., Podtelejnikov, A. V., Molina, H., Bickel, P. E., Andersen, J. S., Fernandez, M. M., Bunkenborg, J., Roepstorff, P., Kristiansen, K., Lodish, H. F., Mann, M., & Pandey, A. (2002). A proteomic approach for identification of secreted proteins during the differentiation of 3T3-L1 preadipocytes to adipocytes. Molecular and Cellular Proteomics, 1(3), 213-22.

Kratchmarova, I, Kalume, DE, Blagoev, B, Scherer, PE, Podtelejnikov, AV, Molina, H, Bickel, PE, Andersen, JS, Fernandez, MM, Bunkenborg, J, Roepstorff, P, Kristiansen, K, Lodish, HF, Mann, M & Pandey, A 2002, 'A proteomic approach for identification of secreted proteins during the differentiation of 3T3-L1 preadipocytes to adipocytes', Molecular and Cellular Proteomics, vol. 1, no. 3, pp. 213-22.

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title = "A proteomic approach for identification of secreted proteins during the differentiation of 3T3-L1 preadipocytes to adipocytes",

abstract = "We have undertaken a systematic proteomic approach to purify and identify secreted factors that are differentially expressed in preadipocytes versus adipocytes. Using one-dimensional gel electrophoresis combined with nanoelectrospray tandem mass spectrometry, proteins that were specifically secreted by 3T3-L1 preadipocytes or adipocytes were identified. In addition to a number of previously reported molecules that are up- or down-regulated during this differentiation process (adipsin, adipocyte complement-related protein 30 kDa, complement C3, and fibronectin), we identified four secreted molecules that have not been shown previously to be expressed differentially during the process of adipogenesis. Pigment epithelium-derived factor, a soluble molecule with potent antiangiogenic properties, was found to be highly secreted by preadipocytes but not adipocytes. Conversely, we found hippocampal cholinergic neurostimulating peptide, neutrophil gelatinase-associated lipocalin, and haptoglobin to be expressed highly by mature adipocytes. We also used liquid chromatography-based separation followed by automated tandem mass spectrometry to identify proteins secreted by mature adipocytes. Several additional secreted proteins including resistin, secreted acidic cysteine-rich glycoprotein/osteonectin, stromal cell-derived factor-1, cystatin C, gelsolin, and matrix metalloprotease-2 were identified by this method. To our knowledge, this is the first study to identify several novel secreted proteins by adipocytes by a proteomic approach using mass spectrometry.",

author = "Irina Kratchmarova and Kalume, {Dario E} and Blagoy Blagoev and Scherer, {Philipp E} and Podtelejnikov, {Alexandre V} and Henrik Molina and Bickel, {Perry E} and Andersen, {Jens S} and Fernandez, {Minerva M} and Jacob Bunkenborg and Peter Roepstorff and Karsten Kristiansen and Lodish, {Harvey F} and Matthias Mann and Akhilesh Pandey",

note = "Keywords: 3T3 Cells; Acute-Phase Proteins; Adipocytes; Animals; Cell Differentiation; Eye Proteins; Gene Expression; Haptoglobins; Lipocalins; Mice; Nerve Growth Factors; Neuropeptides; Oncogene Proteins; Proteins; Proteome; Serpins; Spectrometry, Mass, Electrospray Ionization",

year = "2002",

language = "English",

volume = "1",

pages = "213--22",

journal = "Molecular and Cellular Proteomics",

issn = "1535-9476",

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T1 - A proteomic approach for identification of secreted proteins during the differentiation of 3T3-L1 preadipocytes to adipocytes

AU - Kratchmarova, Irina

AU - Kalume, Dario E

AU - Blagoev, Blagoy

AU - Scherer, Philipp E

AU - Podtelejnikov, Alexandre V

AU - Molina, Henrik

AU - Bickel, Perry E

AU - Andersen, Jens S

AU - Fernandez, Minerva M

AU - Bunkenborg, Jacob

AU - Roepstorff, Peter

AU - Kristiansen, Karsten

AU - Lodish, Harvey F

AU - Mann, Matthias

AU - Pandey, Akhilesh

N1 - Keywords: 3T3 Cells; Acute-Phase Proteins; Adipocytes; Animals; Cell Differentiation; Eye Proteins; Gene Expression; Haptoglobins; Lipocalins; Mice; Nerve Growth Factors; Neuropeptides; Oncogene Proteins; Proteins; Proteome; Serpins; Spectrometry, Mass, Electrospray Ionization

PY - 2002

Y1 - 2002

N2 - We have undertaken a systematic proteomic approach to purify and identify secreted factors that are differentially expressed in preadipocytes versus adipocytes. Using one-dimensional gel electrophoresis combined with nanoelectrospray tandem mass spectrometry, proteins that were specifically secreted by 3T3-L1 preadipocytes or adipocytes were identified. In addition to a number of previously reported molecules that are up- or down-regulated during this differentiation process (adipsin, adipocyte complement-related protein 30 kDa, complement C3, and fibronectin), we identified four secreted molecules that have not been shown previously to be expressed differentially during the process of adipogenesis. Pigment epithelium-derived factor, a soluble molecule with potent antiangiogenic properties, was found to be highly secreted by preadipocytes but not adipocytes. Conversely, we found hippocampal cholinergic neurostimulating peptide, neutrophil gelatinase-associated lipocalin, and haptoglobin to be expressed highly by mature adipocytes. We also used liquid chromatography-based separation followed by automated tandem mass spectrometry to identify proteins secreted by mature adipocytes. Several additional secreted proteins including resistin, secreted acidic cysteine-rich glycoprotein/osteonectin, stromal cell-derived factor-1, cystatin C, gelsolin, and matrix metalloprotease-2 were identified by this method. To our knowledge, this is the first study to identify several novel secreted proteins by adipocytes by a proteomic approach using mass spectrometry.

AB - We have undertaken a systematic proteomic approach to purify and identify secreted factors that are differentially expressed in preadipocytes versus adipocytes. Using one-dimensional gel electrophoresis combined with nanoelectrospray tandem mass spectrometry, proteins that were specifically secreted by 3T3-L1 preadipocytes or adipocytes were identified. In addition to a number of previously reported molecules that are up- or down-regulated during this differentiation process (adipsin, adipocyte complement-related protein 30 kDa, complement C3, and fibronectin), we identified four secreted molecules that have not been shown previously to be expressed differentially during the process of adipogenesis. Pigment epithelium-derived factor, a soluble molecule with potent antiangiogenic properties, was found to be highly secreted by preadipocytes but not adipocytes. Conversely, we found hippocampal cholinergic neurostimulating peptide, neutrophil gelatinase-associated lipocalin, and haptoglobin to be expressed highly by mature adipocytes. We also used liquid chromatography-based separation followed by automated tandem mass spectrometry to identify proteins secreted by mature adipocytes. Several additional secreted proteins including resistin, secreted acidic cysteine-rich glycoprotein/osteonectin, stromal cell-derived factor-1, cystatin C, gelsolin, and matrix metalloprotease-2 were identified by this method. To our knowledge, this is the first study to identify several novel secreted proteins by adipocytes by a proteomic approach using mass spectrometry.

M3 - Journal article

C2 - 12096121

SN - 1535-9476

VL - 1

SP - 213

EP - 222

JO - Molecular and Cellular Proteomics

JF - Molecular and Cellular Proteomics

IS - 3

ER -

A proteomic approach for identification of secreted proteins during the differentiation of 3T3-L1 preadipocytes to adipocytes

Abstract

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