Proteomic identification of matrix metalloproteinase substrates in the human vasculature

Christin Stegemann, Athanasios Didangelos, Javier Barallobre-Barreiro, Sarah R. Langley, Kaushik Mandal, Marjan Jahangiri, Manuel Mayr

Research output: Contribution to journalReview article

32 Citations (Scopus)

Abstract

Background-Matrix metalloproteinases (MMPs) play a key role in cardiovascular disease, in particular aneurysm formation and plaque rupture. Surprisingly, little is known about MMP substrates in the vasculature. Methods and Results-We used a proteomics approach to identify vascular substrates for 3 MMPs, 1 of each of the 3 major classes of MMPs: Human arteries were incubated with MMP-3 (a member of stromelysins), MMP-9 (considered a gelatinase), and MMP-14 (considered a member of the collagenases and of the membrane-bound MMPs). Candidate substrates were identified by mass spectrometry based on increased release from the arterial tissue on digestion, spectral evidence for proteolytic degradation after gel separation, and identification of nontryptic cleavage sites. Using this approach, novel candidates were identified, including extracellular matrix proteins associated with the basement membrane, elastic fibers (emilin-1), and other extracellular proteins (periostin, tenascin-X). Seventy-four nontryptic cleavage sites were detected, many of which were shared among different MMPs. The proteomics findings were validated by immunoblotting and by digesting recombinant/purified proteins with exogenous MMPs. As proof-of-principle, results were related to in vivo pathology by searching for corresponding degradation products in human aortic tissue with different levels of endogenous MMP-9. Conclusions-The application of proteomics to identify MMP targets is a new frontier in cardiovascular research. Our current classification of MMPs based on few substrates is an oversimplification of a complex area of biology. This study provides a more comprehensive assessment of potential MMP substrates in the vasculature and represents a valuable resource for future investigations.

Original languageEnglish (US)
Pages (from-to)106-117
Number of pages12
JournalCirculation: Cardiovascular Genetics
Volume6
Issue number1
DOIs
StatePublished - Feb 1 2013

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Matrix Metalloproteinases
Proteomics
Matrix Metalloproteinase 3
Matrix Metalloproteinase 9
Matrix Metalloproteinase 14
Gelatinases
Matrix Metalloproteinase 1
Elastic Tissue
Collagenases
Immunoblotting
Basement Membrane
Aneurysm
Extracellular Matrix
Blood Vessels
Rupture
Digestion
Mass Spectrometry
Cardiovascular Diseases
Arteries
Gels

All Science Journal Classification (ASJC) codes

  • Genetics
  • Cardiology and Cardiovascular Medicine
  • Genetics(clinical)

Cite this

Stegemann, C., Didangelos, A., Barallobre-Barreiro, J., Langley, S. R., Mandal, K., Jahangiri, M., & Mayr, M. (2013). Proteomic identification of matrix metalloproteinase substrates in the human vasculature. Circulation: Cardiovascular Genetics, 6(1), 106-117. https://doi.org/10.1161/CIRCGENETICS.112.964452
Stegemann, Christin ; Didangelos, Athanasios ; Barallobre-Barreiro, Javier ; Langley, Sarah R. ; Mandal, Kaushik ; Jahangiri, Marjan ; Mayr, Manuel. / Proteomic identification of matrix metalloproteinase substrates in the human vasculature. In: Circulation: Cardiovascular Genetics. 2013 ; Vol. 6, No. 1. pp. 106-117.
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Stegemann, C, Didangelos, A, Barallobre-Barreiro, J, Langley, SR, Mandal, K, Jahangiri, M & Mayr, M 2013, 'Proteomic identification of matrix metalloproteinase substrates in the human vasculature', Circulation: Cardiovascular Genetics, vol. 6, no. 1, pp. 106-117. https://doi.org/10.1161/CIRCGENETICS.112.964452

Proteomic identification of matrix metalloproteinase substrates in the human vasculature. / Stegemann, Christin; Didangelos, Athanasios; Barallobre-Barreiro, Javier; Langley, Sarah R.; Mandal, Kaushik; Jahangiri, Marjan; Mayr, Manuel.

In: Circulation: Cardiovascular Genetics, Vol. 6, No. 1, 01.02.2013, p. 106-117.

Research output: Contribution to journalReview article

TY - JOUR

T1 - Proteomic identification of matrix metalloproteinase substrates in the human vasculature

AU - Stegemann, Christin

AU - Didangelos, Athanasios

AU - Barallobre-Barreiro, Javier

AU - Langley, Sarah R.

AU - Mandal, Kaushik

AU - Jahangiri, Marjan

AU - Mayr, Manuel

PY - 2013/2/1

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N2 - Background-Matrix metalloproteinases (MMPs) play a key role in cardiovascular disease, in particular aneurysm formation and plaque rupture. Surprisingly, little is known about MMP substrates in the vasculature. Methods and Results-We used a proteomics approach to identify vascular substrates for 3 MMPs, 1 of each of the 3 major classes of MMPs: Human arteries were incubated with MMP-3 (a member of stromelysins), MMP-9 (considered a gelatinase), and MMP-14 (considered a member of the collagenases and of the membrane-bound MMPs). Candidate substrates were identified by mass spectrometry based on increased release from the arterial tissue on digestion, spectral evidence for proteolytic degradation after gel separation, and identification of nontryptic cleavage sites. Using this approach, novel candidates were identified, including extracellular matrix proteins associated with the basement membrane, elastic fibers (emilin-1), and other extracellular proteins (periostin, tenascin-X). Seventy-four nontryptic cleavage sites were detected, many of which were shared among different MMPs. The proteomics findings were validated by immunoblotting and by digesting recombinant/purified proteins with exogenous MMPs. As proof-of-principle, results were related to in vivo pathology by searching for corresponding degradation products in human aortic tissue with different levels of endogenous MMP-9. Conclusions-The application of proteomics to identify MMP targets is a new frontier in cardiovascular research. Our current classification of MMPs based on few substrates is an oversimplification of a complex area of biology. This study provides a more comprehensive assessment of potential MMP substrates in the vasculature and represents a valuable resource for future investigations.

AB - Background-Matrix metalloproteinases (MMPs) play a key role in cardiovascular disease, in particular aneurysm formation and plaque rupture. Surprisingly, little is known about MMP substrates in the vasculature. Methods and Results-We used a proteomics approach to identify vascular substrates for 3 MMPs, 1 of each of the 3 major classes of MMPs: Human arteries were incubated with MMP-3 (a member of stromelysins), MMP-9 (considered a gelatinase), and MMP-14 (considered a member of the collagenases and of the membrane-bound MMPs). Candidate substrates were identified by mass spectrometry based on increased release from the arterial tissue on digestion, spectral evidence for proteolytic degradation after gel separation, and identification of nontryptic cleavage sites. Using this approach, novel candidates were identified, including extracellular matrix proteins associated with the basement membrane, elastic fibers (emilin-1), and other extracellular proteins (periostin, tenascin-X). Seventy-four nontryptic cleavage sites were detected, many of which were shared among different MMPs. The proteomics findings were validated by immunoblotting and by digesting recombinant/purified proteins with exogenous MMPs. As proof-of-principle, results were related to in vivo pathology by searching for corresponding degradation products in human aortic tissue with different levels of endogenous MMP-9. Conclusions-The application of proteomics to identify MMP targets is a new frontier in cardiovascular research. Our current classification of MMPs based on few substrates is an oversimplification of a complex area of biology. This study provides a more comprehensive assessment of potential MMP substrates in the vasculature and represents a valuable resource for future investigations.

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DO - 10.1161/CIRCGENETICS.112.964452

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