Immunolocalization of tissue inhibitor of metalloproteinases (TIMP) in human cells. Characterization and use of a specific antiserum

A specific antiserum to pure human amniotic fluid metalloproteinase inhibitor (TIMP) was raised in a sheep. This antiserum was used to demonstrate: firstly, the immunological identity of the TIMP activities from amniotic fluid and culture medium of human foetal lung fibroblasts; and secondly, by indirect immunofluorescence, the secretion of TIMP by human foetal lung fibroblasts, chondrocytes, epithelial cells and smooth muscle cells. The phorbol ester, 12-O-tetradecanoylphorbol-13-acetate, was used to stimulate secretion of TIMP by human foetal lung fibroblasts and the ionophore monensin was used to demonstrate intracellular accumulation of TIMP in the Golgi apparatus of these cells. These results are discussed in relation to other inhibitors of collagenase reported in the literature, which are probably identical to TIMP.

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Large inhibitor of metalloproteinases (LIMP) contains tissue inhibitor of metalloproteinases (TIMP)-2 bound to 72,000-Mr progelatinase

Biochemical Journal ◽

10.1042/bj2850143 ◽

1992 ◽

Vol 285 (1) ◽

pp. 143-147 ◽

Cited By ~ 12

Author(s):

V A Curry ◽

I M Clark ◽

H Bigg ◽

T E Cawston

Keyword(s):

Extracellular Matrix ◽

Connective Tissue ◽

Inhibitory Activity ◽

Culture Medium ◽

Lung Fibroblasts ◽

Tissue Inhibitor Of Metalloproteinases ◽

Tissue Inhibitor ◽

Metalloproteinase Inhibitor ◽

Cells In Culture ◽

Foetal Lung

Connective-tissue cells in culture produce a family of metalloproteinases which, once activated, can degrade all the components of the extracellular matrix. These potent enzymes are all inhibited by the tissue inhibitor of metalloproteinases (TIMP), and it was thought that this inhibitor was solely responsible for the inhibition of these enzymes within connective tissue. However, other inhibitors have recently been described, including large inhibitor of metalloproteinases (LIMP) present in the culture medium of human foetal lung fibroblasts. Here we show that a large proportion of the inhibitory activity of LIMP consists of 72,000-M(r)-progelatinase bound to TIMP-2, a recently discovered low-M(r) metalloproteinase inhibitor closely related to TIMP. The physiological implications of the secretion of a complex of 72,000-M(r) progelatinase and TIMP-2 are discussed, and the separation of the complex in 6 M-urea is described.

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Rapid purification of tissue inhibitor of metalloproteinases from human plasma and identification as a γ-serum protein

Biochemical Journal ◽

10.1042/bj2380677 ◽

1986 ◽

Vol 238 (3) ◽

pp. 677-682 ◽

Cited By ~ 17

Author(s):

T E Cawston ◽

D N Noble ◽

G Murphy ◽

A J Smith ◽

C Woodley ◽

...

Keyword(s):

Synovial Fluid ◽

Amniotic Fluid ◽

Culture Medium ◽

Gel Filtration ◽

Tissue Inhibitor Of Metalloproteinases ◽

Human Amniotic Fluid ◽

Tissue Inhibitor ◽

Serum Component ◽

Rapid Purification ◽

Human Synovial Fluid

A rapid method is described for the purification of human tissue inhibitor of metalloproteinases (TIMP) from plasma which involves immuno-affinity chromatography and gel filtration. The purified plasma inhibitor is immunologically identical with the TIMP previously purified from human amniotic fluid, human synovial fluid and human fibroblast culture medium. It is proposed that this inhibitor is identical with the plasma inhibitor previously named ‘B1 anticollagenase’, although the plasma inhibitor was shown to migrate as a gamma-serum component.

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Cell-free human amniotic fluid as culture medium for mouse and human embryos

Fertility and Sterility ◽

10.1016/s0015-0282(16)60619-2 ◽

1989 ◽

Vol 51 (4) ◽

pp. 671-674 ◽

Cited By ~ 6

Author(s):

Andrew D. Dorfmann ◽

Samuel D. Bender ◽

Patricia Robinson ◽

Edward F. Fugger ◽

Maria Bustillo ◽

...

Keyword(s):

Amniotic Fluid ◽

Culture Medium ◽

Human Embryos ◽

Human Amniotic Fluid

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Characterization of a specific antiserum for mammalian collagenase from several species: immunolocalization of collagenase in rabbit chondrocytes and uterus

Journal of Cell Science ◽

10.1242/jcs.81.1.105 ◽

1986 ◽

Vol 81 (1) ◽

pp. 105-123 ◽

Cited By ~ 5

Author(s):

R.M. Hembry ◽

G. Murphy ◽

T.E. Cawston ◽

J.T. Dingle ◽

J.J. Reynolds

Keyword(s):

Post Partum ◽

Mammalian Species ◽

Specific Antiserum ◽

Gravid Uterus ◽

Intracellular Accumulation ◽

Normal Cartilage ◽

Active Collagenase ◽

Rabbit Bone ◽

Ionophore Monensin

A specific antiserum to rabbit bone collagenase was raised in a sheep and shown to react with collagenase from several mammalian species. This antiserum was used to demonstrate that only active collagenase binds to and can be immunolocalized on collagen fibrils and, by indirect immunofluorescence, the secretion of latent collagenase by stimulated rabbit chondrocytes and cells of the post-partum involuting rabbit uterus. The ionophore monensin was used to demonstrate intracellular accumulation of collagenase in the Golgi apparatus of both stimulated chondrocytes and involuting uterine cells. Collagenase was not detectable in either normal cartilage or non-gravid uterus. These results are discussed in relation to other studies of collagenase immunolocalization reported in the literature.

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Influences of endogenous and exogenous TGF-beta on elastin in rat lung fibroblasts and aortic smooth muscle cells

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1992.263.2.l257 ◽

1992 ◽

Vol 263 (2) ◽

pp. L257-L263 ◽

Cited By ~ 6

Author(s):

S. E. McGowan

Keyword(s):

Smooth Muscle ◽

Steady State ◽

Smooth Muscle Cells ◽

Culture Medium ◽

Muscle Cells ◽

Neonatal Rat ◽

Lung Fibroblasts ◽

Rat Lung ◽

Tgf Beta ◽

Adult Rats

The factors that regulate elastin production during neonatal lung development have not been elucidated. Previous investigations suggested that transforming growth factor-beta (TGF-beta) increases elastin production by neonatal rat lung fibroblasts (LF). We examined whether this effect of TGF-beta was unique to these cells or was evident in other neonatal cells, which constitutively produce elastin, or in cells from adults, whose constitutive elastin production is low. We have quantitated soluble elastin, elastin mRNA, and TGF-beta production in primary cultures of smooth muscle cells (SMC) and LF from neonatal and adult rats and have examined the alterations in soluble elastin and elastin mRNA that result from adding 100 pM exogenous TGF-beta 1 to these cultures. Unsupplemented cultures of LF and SMC obtained from neonatal rats exhibited higher steady-state levels of elastin mRNA and contained more soluble elastin in their culture medium than did cells from adult animals. When neonatal LF were supplemented with 100 pM TGF-beta 1, they showed a significant increase in the soluble elastin content of their culture medium and their steady-state elastin mRNA. Neither LF obtained from adults nor SMC obtained from neonatal or adult rats significantly increased their soluble elastin or steady-state elastin mRNA after the addition of exogenous TGF-beta. When neonatal LF were supplemented with an anti-TGF-beta neutralizing antibody, the soluble elastin content of the culture medium decreased significantly. These data suggest that the responsiveness of elastin expression to TGF-beta is limited to neonatal LF and that endogenous TGF-beta influences elastin production by neonatal LF.

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C67 SMOOTH MUSCLE CELLS DIFFERENTIATED FROM HUMAN AMNIOTIC FLUID DERIVED STEM CELLS AS A POSSIBLE TREATMENT OF BLADDER DYSFUNCTION

European Urology Supplements ◽

10.1016/s1569-9056(13)60067-6 ◽

2012 ◽

Vol 11 (4) ◽

pp. 97

Author(s):

J. Olkowska ◽

N. Gurtowska ◽

A. Bajek ◽

M. Walentowicz-Sadlecka ◽

P. Sadlecki ◽

...

Keyword(s):

Stem Cells ◽

Smooth Muscle ◽

Smooth Muscle Cells ◽

Amniotic Fluid ◽

Bladder Dysfunction ◽

Muscle Cells ◽

Human Amniotic Fluid

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An inhibitor of collagenase from human amniotic fluid. Purification, characterization and action on metalloproteinases

Biochemical Journal ◽

10.1042/bj1950167 ◽

1981 ◽

Vol 195 (1) ◽

pp. 167-170 ◽

Cited By ~ 103

Author(s):

G Murphy ◽

T E Cawston ◽

J J Reynolds

Keyword(s):

Amniotic Fluid ◽

Tissue Inhibitor Of Metalloproteinases ◽

Human Amniotic Fluid ◽

Tissue Inhibitor ◽

Number Of Species

1. An inhibitor of collagenase of apparent mol.wt. 28000 was isolated from term human amniotic fluid. 2. It is active against mammalian collagenases from a number of species and tissues as well as other mammalian metalloproteinases, but has no activity against bacterial metalloproteinases. 3. Activity is destroyed by treatment with either trypsin or 4-aminophenylmercuric acetate, by heat, and by reduction and carboxymethylation. 4. All the properties observed suggest that it is similar to the synthesized tissue inhibitor of metalloproteinases.

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