Structural analysis of tissue inhibitor of metalloproteinases-1 (TIMP-1) by tryptic peptide mapping

doi:10.1016/0167-4838(93)90105-Z

Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology

Volume 1164, Issue 1, 24 June 1993, Pages 8-16

Biochimica et Biophy...

https://doi.org/10.1016/0167-4838(93)90105-Z Get rights and content

Abstract

Tryptic digests of recombinant TIMP-1 have been resolved on reverse-phase HPLC and the major peaks identified by N-terminal sequencing. This procedure accounted for the entire molecule, except two short peptides of 2 and 4 amino acids in length. The peptide map was used to (i), characterize an insoluble ‘core’ peptide seen on digestion of TIMP-1 in non-reducing conditions; (ii), confirm the structure of Δ_127–184TIMP-1, a recently described truncated form of the TIMP-1 molecule; (iii), identify exposed regions of the intact and truncated TIMP-1 molecules by measuring the rate of tryptic peptide release and (iv), locate sites of aberrant proteolysis seen when recombinant human TIMP-1 was purified at large scale.

References (21)

L.M. Matrisian
Trends Genet.
(1990)
L.A. Liotta et al.
Cell
(1991)
W.G. Stetler-Stevenson et al.
J. Biol. Chem.
(1989)
G. Murphy et al.
Biochim. Biophys. Acta.
(1985)
G.P. Stricklin
Collagen Rel. Res.
(1986)
G.P. Stricklin et al.
J. Biol. Chem.
(1983)
S. Cooksley et al.
Matrix
(1990)
U.K. Laemmli et al.
J. Mol. Biol.
(1973)
G. Murphy et al.
BioEssays
(1985)
T.E. Cawston

There are more references available in the full text version of this article.

Cited by (17)

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The tissue inhibitor of metalloproteinases-1 (TIMP-1) protein can regulate the expression of certain proteases and microRNAs in cancer cells, and it is highly possible to diagnose cancers through analyzing the expression of TIMP-1 on exosomes. However, it is still a great challenge to obtain reliable physiological information on TIMP-1 by label-free method from exosomes in plasma. Here, we designed a porous-plasmonic SERS chip functionalized with synthesized CP05 polypeptide, which can specifically capture and distinguish exosomes from diverse origins. The SERS chip can accurately locate the plasmon in TIMP-1 protein to analyze the discrepancy of related fingerprint peaks of different exosomes. Based on the designed SERS chip, we successfully distinguished the lung and colon cancer cell-derived exosomes from normal exosomes at the single vesicle level by unique Raman spectroscopy and machine learning methods. This work not only provides a practical SERS chip for the application of Raman technology in human tumor monitoring and prognosis, but also provides a new idea for analyzing the feature of exosomes at the spectral level.
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Over the last few years a growing number of matrix degrading metalloproteinases have been implicated in cancer. These include in particular the matrix metalloproteinases (MMPs) which have been shown to be associated with a large variety of human malignancies, and the metalloproteinases with a disintegrin domain (ADAMs) whose potential role in cancer has begun to be examined. The expression of MMPs in human cancer is the result of a complex interaction between tumour cells and non-malignant stromal cells including fibroblasts, endothelial cells and inflammatory cells which all actively participate in the production of MMPs in tumour tissue. The proteolytic modification of the extracellular matrix by these proteases does more than allow malignant cells to locally invade and form distant metastasis. It significantly alters the tumour micro-environment and modifies the contacts between tumour cells and extracellular matrix proteins. These changes can affect essential cellular functions such as growth, survival, migration and even drug resistance. As our understanding of the nature of the contacts between tumour cells and a proteolytically modified extracellular matrix continues to progress, it is likely that novel therapeutic approaches to modify tumour cell behaviour will be identified.
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Chemical modification of tissue inhibitor of metalloproteinases-1 and its inactivation by diethyl pyrocarbonate
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Tissue inhibitor of metalloproteinases-1 (TIMP-1) was treated with a range of chemical modification reagents in order to identify amino acid residues essential for inhibitory activity. Diethyl pyrocarbonate (DEPC) was found to be a potent inactivator at low reagent/TIMP molar concentrations. The extent of modification at 50% inactivation was determined as 1.5 sites/molecule. The DEPC-modified inhibitor did not form stable complexes with stromelysin, but was shown to retain native structure as judged by conformational stability to denaturation by guanidine hydrochloride. Peptide mapping experiments were used to find the sites of DEPC incorporation within the primary structure of TIMP and three residues were identified (His-95, His-144 and His-164). Mutant TIMPs in which histidine residues have been substituted or deleted retain inhibitory activity and were found to be equally as sensitive to DEPC inactivation as the wild-type. No new sites of DEPC modification in the mutant proteins were detected. The possible contribution made by His residues 95, 144 and 164 to the inhibitory activity of TIMP is discussed.
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Structural analysis of tissue inhibitor of metalloproteinases-1 (TIMP-1) by tryptic peptide mapping

Abstract

Trends Genet.

Cell

J. Biol. Chem.

Biochim. Biophys. Acta.

Collagen Rel. Res.

J. Biol. Chem.

Matrix

J. Mol. Biol.

BioEssays