Mammalian collagenases cleave all three polypeptide chains of the triple helical collagen molecule at a specific site to give characteristic one-quarter and three-quarter fragments. These denature at 37�C becoming susceptible to digestion by less specific proteinases.
Zymography and reverse zymography are techniques used to analyze the activities of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in complex biological samples. The two methods are technically similar. Zymography involves the electr ...
The techniques of Western blotting and immunocytochemistry are suitable for the quantification and localization, respectively, of matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) expression. However, they do not indicate the endogenous ba ...
How do cells degrade their surrounding matrix? Constitutive and induced cellular secretion of several classes of proteinases have been implicated in extracellular degradation (1) and many of these proteinases have the ability to degrade extracellular matrix proteins in vitro. Ho ...
The matrix metalloproteinases (MMPs) have a pivotal role in both normal and pathological turnover of the extracellular matrix. Whereas MMP protein can easily be detected by immunolocalization or Western blot analysis, the determination of whether or not an MMP is active and acting on a part ...
Collagen is the most abundant protein in the mammalian body. Collagen types I, II, and III are the major structural components of skin, bone, cartilage, and connective tissues. They exist as fibrils of crosslinked helical molecules composed of three α chains of approx 1000 amino acids each, with non ...
The proteoglycans are a group of macromolecules characterized by the covalent attachment of one or more sulfated glycosaminoglycan (GAG) chains to a protein core, and are an important constituent of the extracellular matrix. Cartilage contains a number of proteoglycans, the most abun ...
Degradation of fibrillar collagens by matrix metalloproteinases (MMPs) is thought to be a major catabolic pathway in various connective tissues. However, the complex structure of collagen molecules and their degradation products makes specific assay of these events very challe ...
Matrix metalloproteinases are believed to play a major role in cellular invasion and tumor metastasis. Therefore, to understand the molecular mechanisms underlying invasion, a variety of in vitro assays have been developed. Most assays assess the interaction of tumor cells with the sur ...
Catabolism of extracellular matrix (ECM) components has been ascribed to a family of Zn2� metalloenzymes. These matrix metalloproteinases (MMPs; also termed matrixins) are believed to be important in connective tissue remodeling during development and wound healing. MMPs have al ...
Tissue inhibitors of metalloproteinases (TIMPs) are well known as inhibitors of matrix metalloproteinases (MMPs) such as interstitial collagenase, gelatinase A and B, and stromelysin 1, and have been suggested to play an important role in the regulation of MMPs. Although MMP inhibition ...
The kinetic analysis of the inhibition of matrix metalloproteinases by tissue inhibitor of metalloproteinases (TIMP) yields valuable information on the mechanism and specificity of the TIMPs. When combined with the use of genetic engineering or chemical methods of modification to ...
The emergence of cycle sequencing (1) as a powerful alternative to conventional isothermal methods has facilitated the manipulations involved in sequencing protocols in general, and is of value for the sequence analysis of PCR products, in particular. Using radioactive labeling tec ...
At least 19 proteins have now been defined as collagens (1,2), but many of those recently discovered are present in tissues in such small amounts that their isolation for characterization at the protein level has so far been impossible. Some of the fibril-forming collagens are now in medical use, in ap ...
Techniques for direct sequencing of PCR products are of central importance to contemporary research in molecular biology and genetics. The rapidly growing number of cloned human disease genes increasingly allows sequencing of PCR amplicons for diagnostic purposes. Nonradioac ...
The conventional methods of sequencing, such as Maxam and Gilbert (1) (involving chemical cleavage of labeled DNA fragments) and Sanger et al. (2), of dideoxy sequencing (using enzymatic extension of oligonucleotide primers) remain the most reliable techniques, but when compared to more ...
The polymerase chain reaction (PCR) allows the rapid detection of infectious viruses or other microorganisms as well as the cause of genetic defects. By performing sequence analysis afterward, important additional information on the PCR products is obtained. Often direct sequenci ...
The polymerase chain reaction (PCR) has become widely established as a powerful core molecular biology technique because of its ability of produce large amounts of specific target DNA from limited template sources (1). Numerous applications based on the PCR have also been developed, incl ...
Among the many techniques of cloning new genes, one approach involves degenerate primers (1–7). The approach usually requires three steps: 1. Using degenerate primers to amplify part of the gene of interest by PCR: The degenerate primers’ sequence
The inherent problems of sensitivity and specificity that one encounters when trying to determine a particular nucleotide sequence directly in its genomic context can be overcome by selective amplification of the region of interest. This amplification of the target DNA is usually ach ...
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