Detection of TIMP-2-like protein in Atlantic cod ( Gadus morhua) muscle using two-dimensional real-time reverse zymography

Matrix metalloproteinases (MMPs) have been proposed to participate in postmortem degradation of fish muscle connective tissues during storage. In the extracellular matrix (ECM) of mammals, a group of specific tissue inhibitors of metalloproteinases (TIMPs) contributes in regulating the MMPs present....

Full description

Saved in:
Bibliographic Details
Published in:Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 2004-10, Vol.139 (2), p.253-259
Main Authors: Lødemel, Jørgen B., Egge-Jacobsen, Wolfgang, Olsen, Ragnar L.
Format: Article
Language:English
Subjects:
2D real-time reverse zymography
Amino Acid Sequence
amino acid sequences
Animals
Autolysis
Cod
Gadus morhua
Gelatinases - antagonists & inhibitors
Humans
Marine
metalloproteinases
Muscle
muscle tissues
Muscles - chemistry
Peptide Fragments
proteinase inhibitors
Sequence Alignment
TIMP
Tissue Inhibitor of Metalloproteinase-2 - analysis
Tissue Inhibitor of Metalloproteinase-2 - isolation & purification
Tissue Inhibitor of Metalloproteinase-2 - pharmacology
Tissue inhibitors of metalloproteinases
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Matrix metalloproteinases (MMPs) have been proposed to participate in postmortem degradation of fish muscle connective tissues during storage. In the extracellular matrix (ECM) of mammals, a group of specific tissue inhibitors of metalloproteinases (TIMPs) contributes in regulating the MMPs present. However, little information exists on the presence of TIMPs in fish. In this paper, the presence of TIMPs in the muscle of Atlantic cod ( Gadus morhua) was investigated using gelatin affinity chromatography, real-time reverse zymography (RTRZ) and mass spectrometry (MS). Using RTRZ inhibitory action against cod muscle, proteinases binding to gelatin were detected in the muscle. The inhibitor had similar molecular weight (21 kDa) as a human recombinant TIMP-2 used as a reference sample. Because isoforms of TIMP-2 homologues with similar molecular weight have been suggested in fish, a two-dimensional RTRZ (2D RTRZ) method was designed. The new method showed the existence of only one form with inhibitory action against cod muscle proteinases. Finally, de novo sequencing of two peptides derived from the cod muscle inhibitor showed high homology to TIMP-2s both from human and other teleosts.
ISSN:1096-4959
1095-6433
1879-1107
DOI:10.1016/j.cbpc.2004.08.004