scholarly journals Expression of human recombinant 72 kDa gelatinase and tissue inhibitor of metalloproteinase-2 (TIMP-2): characterization of complex and free enzyme

1993 ◽  
Vol 289 (2) ◽  
pp. 411-416 ◽  
Author(s):  
R Fridman ◽  
R E Bird ◽  
M Hoyhtya ◽  
M Oelkuct ◽  
D Komarek ◽  
...  
Keyword(s):  
Specific Activity ◽  
Expression System ◽  
Enzymic Activity ◽  
Free Enzyme ◽  
Tissue Inhibitor Of Metalloproteinase ◽  
Molar Ratio ◽  
Collagen Type Iv ◽  
Tissue Inhibitor ◽  
Type Iv Collagenase ◽  
Type Iv

The human 72 kDa gelatinase/type IV collagenase is a metalloproteinase that is thought to play a role in metastasis and angiogenesis. The 72 kDa progelatinase can be isolated from conditioned media as a complex with the tissue inhibitor of metalloproteinase-2 (TIMP-2). To investigate 72 kDa gelatinase-TIMP-2 interactions and to compare the activity of the complex versus that of the free enzyme, we have expressed and purified human 72 kDa progelatinase and TIMP-2 as single proteins in a recombinant vaccinia virus mammalian cell expression system. The recombinant 72 kDa progelatinase was able to bind TIMP-2, and it digested gelatin and collagen type IV after activation by p-aminophenylmercuric acid (APMA). The specific activity of the recombinant free enzyme was 20-fold higher than the activity of an APMA-treated stoichiometric complex of recombinant 72 kDa progelatinase and TIMP-2. Also, TIMP-2 caused an 86% inhibition of activity when added to the activated enzyme at a 1:1 molar ratio. Activation of the free recombinant 72 kDa progelatinase yielded the 62 kDa species and two fragments of 46 and 35 kDa that cross-reacted with monoclonal antibodies to the 72 kDa proenzyme. TIMP-2 inhibited the conversion of the recombinant proenzyme to the 62 kDa species and the appearance of the 45 and 35 kDa bands. These results suggest that TIMP-2 is not only a potent inhibitor of the activated enzyme but also prevents the generation of low-molecular-mass species and full enzymic activity from the zymogen.

Immunolocalization of Human Gelatinase (Type IV Collagenase, MMP-9) and Tissue Inhibitor of Metalloproteinase 1 in Hailey-Hailey and Darier’s Diseases

Dermatology ◽  
1996 ◽  
Vol 193 (2) ◽  
pp. 110-114 ◽  
Author(s):  
T. Kobayashi ◽  
K. Sakuraoka ◽  
S. Hattori ◽  
H. Hori ◽  
Y. Nagai ◽  
...  
Keyword(s):  
Tissue Inhibitor Of Metalloproteinase ◽  
Tissue Inhibitor ◽  
Type Iv Collagenase ◽  
Type Iv

Changes in concentrations of type IV collagen and tissue inhibitor of metalloproteinase-1 in patients with paraquat poisoning

2001 ◽  
Vol 21 (6) ◽  
pp. 445-447 ◽  
Author(s):  
Tsukasa Nakamura ◽  
Chifuyu Ushiyama ◽  
Noriaki Shimada ◽  
Kayo Hayashi ◽  
Isao Ebihara ◽  
...  
Keyword(s):  
Type Iv Collagen ◽  
Tissue Inhibitor Of Metalloproteinase ◽  
Tissue Inhibitor ◽  
Type Iv ◽  
Paraquat Poisoning

scholarly journals Pericyte recruitment during vasculogenic tube assembly stimulates endothelial basement membrane matrix formation

Blood ◽  
2009 ◽  
Vol 114 (24) ◽  
pp. 5091-5101 ◽  
Author(s):  
Amber N. Stratman ◽  
Kristine M. Malotte ◽  
Rachel D. Mahan ◽  
Michael J. Davis ◽  
George E. Davis
Keyword(s):  
Basement Membrane ◽  
Tube Formation ◽  
Tissue Inhibitor Of Metalloproteinase ◽  
Collagen Type Iv ◽  
Matrix Assembly ◽  
Matrix Deposition ◽  
Vascular Basement Membrane ◽  
Membrane Matrix ◽  
Type Iv ◽  
Endothelial Basement Membrane

AbstractWe show that endothelial cell (EC)–generated vascular guidance tunnels (ie, matrix spaces created during tube formation) serve as conduits for the recruitment and motility of pericytes along EC ablumenal surfaces to facilitate vessel maturation events, including vascular basement membrane matrix assembly and restriction of EC tube diameter. During quail development, pericyte recruitment along microvascular tubes directly correlates with vascular basement membrane matrix deposition. Pericyte recruitment to EC tubes leads to specific induction of fibronectin and nidogen-1 (ie, matrix-bridging proteins that link together basement membrane components) as well as perlecan and laminin isoforms. Coincident with these events, up-regulation of integrins, α5β1, α3β1, α6β1, and α1β1, which bind fibronectin, nidogens, laminin isoforms, and collagen type IV, occurs in EC-pericyte cocultures, but not EC-only cultures. Integrin-blocking antibodies to these receptors, disruption of fibronectin matrix assembly, and small interfering RNA suppression of pericyte tissue inhibitor of metalloproteinase (TIMP)-3 (a known regulator of vascular tube stabilization) all lead to decreased EC basement membrane, resulting in increased vessel lumen diameter, a key indicator of dysfunctional EC-pericyte interactions. Thus, pericyte recruitment to EC-lined tubes during vasculogenesis is a stimulatory event controlling vascular basement membrane matrix assembly, a fundamental maturation step regulating the transition from vascular morphogenesis to stabilization.

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