细胞技术

The matrix metalloproteinase field has a clearly defined starting point, the seminal study of Jerome Gross and Charles Lapi�re in 1962 (1) in which fragments of resorbing tadpole tail were cultured on reconstituted collagen gels. A collagenolytic enzyme was recovered from the culture med ...

Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) play important roles in the remodeling of connective tissues associated with normal mammalian development and growth, and in the degradative processes accompanying diseases such as rhe ...

The proteolytic activity of the matrix metalloproteinases (MMPs) involved in extracellular matrix degradation must be precisely regulated by their endogenous protein inhibitors, the tissue inhibitors of metalloproteinases (TIMPs). Disruption of this balance results in s ...

Developmental and homeostatic remodeling of the extracellular matrix (ECM) is a highly regulated process orchestrated by a family of zinc-containing, calcium-dependent neutral proteases known as the matrix metallo-proteinases (MMP). This family of enzymes, which now contains ...

Connective tissue degradation occurs in chronic inflammatory diseases such as arthritis and periodontitis to adversely affect life quality. More importantly, disturbances in connective tissue homeostasis may be life threatening in various lung, neurological, and cardiov ...

The most powerful approach for studying gene function in an intact animal is to regulate the levels of the gene product and thereby see gains-of-function or losses-of-function. The occasional mutation in the genes for the matrix metalloproteinases or their inhibitors, or polymorphism in t ...

Since the discovery of the membrane-type matrix metalloproteinases (MT-MMPs) which are characterized by the existence of a C-terminal membrane-spanning domain followed by a short cytoplasmic “tail” (1) there has been much interest in the production of recombinant protein to facili ...

Deciding what system to use for recombinant matrix metalloproteinase (MMP) expression is often a matter of practicality (i.e., simplicity, cost effectiveness, time considerations). Of course, the final judgment of practicality is determined by the production of sufficient quant ...

E. coli is a convenient host in which to express recombinant proteins. The technology is available to most laboratories as it is relatively inexpensive and does not require extensive expertise. The major drawback of E. coli as an expression host is the inability of the organism to carry out many postt ...

Structural analysis of any protein, whether by X-ray crystallography or NMR, requires a reliable source of large amounts of good quality protein. Most proteins are not sufficiently abundant in their natural state to be used as the primary source, and so it is essential for recombinant protein to be ...

Currently more than twenty different matrix metalloproteinases (MMPs) and four tissue inhibitors of metalloproteinases (TIMPs) have been identified (1,2) by cDNA cloning. This chapter describes methods to purify collagenases (MMP-1, MMP-8, and MMP-13), gelatinases (MMP-2 and MMP ...

With the advent of recombinant DNA technology, numerous systems have been utilized for the overexpression of proteins. Recombinant protein expression in Escherichia coli (E. coli) typically provides large quantities of the protein of interest in a relatively short period of time. The e ...

Resident cells of tissues are capable of secreting an array of structurally related zinc endopeptidases known as matrix metalloproteinases (MMPs). They initiate the degradation of the surrounding macromolecules of the extracellular matrix (ECM), mostly proteoglycans and spe ...

Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) is a sensitive and rapid method to monitor the expression of specific mRNAs. Here we describe two approaches to quantification of steady-state levels of Matrix Metalloproteinase (MMP) and Tissue Inhibitor of Metallopro ...

Many metalloproteinase genes are transcriptionally regulated by differentiation, growth factors, cytokines, and hormones. Nuclear run-off assays and analysis of heterogeneous nuclear RNA (hnRNA) levels are two methods for studying changes in gene transcription. In this cha ...

The activity of matrix metalloproteinases (MMPs) is closely regulated by tissue inhibitors of metalloproteinases (TIMPs) under pathophysiological conditions. The quantitative imbalance between MMPs and TIMPs in tissues and body fluids is considered to cause tumor invasion, ...

The imminent completion of the human and mouse genome projects will reveal all metalloproteases and metalloprotease inhibitors such as those belonging to the MMP, TIMP, ADAM, and ADAMTS (1) families. At the present time, there are over 65 published genes in these families and it is very likely that t ...

Immunohistochemical techniques are a convenient method to identify the cells responsible for the production of MMPs and TIMPs in local tissues under pathophysiological conditions. Direct and indirect methods are presented for immunohistochemistry, but the latter is usually u ...

The degradation of the extracellular matrix during development and in disease is thought to result from the combined action of several proteolytic enzyme systems, including the matrix metalloproteinases (MMPs), serine proteinases, and cysteine proteinases. The majority of the s ...

The essential role of genes is in encoding structural proteins and enzymes which enable the cell or organism to maintain homeostasis in the face of the environmental challenges experienced (1). DNA containing such genetic information varies from one species to another. Even within a speci ...