A proteomic approach to identify substrates of matrix metalloproteinase-14 in human plasma

In Kwan Hwang, Sung Min Park, Se Yeon Kim, Seung-Taek Lee

Research output: Contribution to journalArticle

72 Citations (Scopus)

Abstract

Matrix metalloproteinases (MMPs) are a family of zinc-containing endopeptidases that proteolyze extracellular matrix components as well as a variety of functional proteins. Here we describe a "degradomics" method that efficiently identifies substrates of MMP-14 in a complex protein mixture, such as plasma. Plasma proteins were incubated in the presence or absence of the MMP-14 catalytic domain and displayed on two-dimensional (2-D) gels. After a comparison of the gels, we selected 40 protein spots that reproducibly showed disparities. Upon in-gel digestion, mass determination, and peptide mass fingerprinting, we identified 15 different proteins from 31 spots. These proteins included six known substrates and nine potential substrates of MMP-14. Among the latter, the purified forms of apolipoprotein A-I, apolipoprotein E, and plasma gelsolin were cleaved in vitro by MMP-14, confirming that each of them is a novel substrate of MMP-14. These results demonstrate that our method rapidly and selectively identifies MMP-14 substrates from human plasma proteins. This method would thus constitute a powerful tool for identifying the substrates of MMPs and other proteases in highly complex mixtures of proteins and would enhance our understanding of the biological roles of these enzymes.

Original languageEnglish
Pages (from-to)79-87
Number of pages9
JournalBiochimica et Biophysica Acta - Proteins and Proteomics
Volume1702
Issue number1
DOIs
Publication statusPublished - 2004 Oct 1

Fingerprint

Matrix Metalloproteinase 14
Plasma (human)
Proteomics
Substrates
Proteins
Gels
Complex Mixtures
Matrix Metalloproteinases
Blood Proteins
Gelsolin
Plasmas
Endopeptidases
Peptide Mapping
Apolipoprotein A-I
Apolipoproteins E
Extracellular Matrix
Zinc
Digestion
Catalytic Domain
Peptide Hydrolases

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biophysics
  • Biochemistry
  • Molecular Biology

Cite this

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A proteomic approach to identify substrates of matrix metalloproteinase-14 in human plasma. / Hwang, In Kwan; Park, Sung Min; Kim, Se Yeon; Lee, Seung-Taek.

In: Biochimica et Biophysica Acta - Proteins and Proteomics, Vol. 1702, No. 1, 01.10.2004, p. 79-87.

Research output: Contribution to journalArticle

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