Role of MMP-2 in oxidant-mediated regulation of Ca²⁺ uptake in microsomes of bovine pulmonary artery smooth muscle
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Title |
Role of MMP-2 in oxidant-mediated regulation of Ca²⁺ uptake in microsomes of bovine pulmonary artery smooth muscle
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Creator |
Mandala, Amritlal
Chakrabortia, Tapati Choudhurya, Rajdeep Ghosha, Biswarup Chakrabortia, Sajal |
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Subject |
Pulmonary artery smooth muscle
microsomes oxidant tert-butylhydroperoxide antioxidant vitamin E matrix metalloproteinase-2 tissue inhibitor of metalloproteinase-2 Ca²⁺-ATPase ATP-dependent Ca²⁺ uptake Na⁺-dependent Ca²⁺ uptake |
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Description |
19-27
Treatment of bovine pulmonary artery smooth muscle microsomes with tert-butylhydroperoxide (t-buOOH) (300 µM) markedly stimulated matrix metalloproteinase-2 (MMP-2) activity and enhanced Ca²⁺-ATPase activity and ATP-dependent Ca²⁺ uptake. Pre-treatment with vit. E (1 mM) and tissue inhibitor of metalloproteinase-2 (TIMP-2) (50 µg/ml) prevented t-buOOH-induced stimulation of MMP-2 activity, Ca²⁺-ATPase activity and ATP-dependent Ca²⁺ uptake. In contrast, Na⁺-dependent Ca²⁺ uptake was inhibited by t-buOOH and the inhibition was reversed by vit. E (1 mM) and TIMP-2 (50 µg/ml). However, t-buOOH-triggered changes in MMP-2 activity, and ATP- and Na+-dependent Ca²⁺ uptake were not reversed upon pre-treatment of the microsomes with a low concentration of 5 µg/ml of TIMP-2, which on the contrary reversed MMP-2 (1 µg/ml)-mediated alteration on these parameters. The inhibition of Na+-dependent Ca²⁺ uptake by MMP-2 under t-buOOH treatment overpowered the stimulation of ATP-dependent Ca²⁺ uptake in the microsomes. Combined treatment of the microsomes with low doses of MMP-2 (0.5 µg/ml) and t-buOOH (100 mM) augmented Ca²⁺-ATPase activity and ATP-dependent Ca²⁺ uptake, but inhibited Na+-dependent Ca²⁺ uptake, compared to that elicited by either MMP-2 (0.5 µg/ml) or t-buOOH (100 µM). Pre-treatment with TIMP-2 (50 µg/ml) reversed the effects of MMP-2 (0.5 µg/ml) and/or t-buOOH (100 mM). Although pre-treatment with 5 µg/ml of TIMP-2 reversed the effects produced by MMP-2 (0.5 µg/ml), but it did not inhibit the responses elicited by t-buOOH (300 µM) or t-buOOH (100 mM) plus MMP-2 (0.5 mg/ml) in the microsomes. Treatment with TIMP-2 (5 mg/ml) inhibited MMP-2 (1 mg/ml) activity (assessed by [14C]-gelatin degradation), whereas treatment of t-buOOH (300 µM) with TIMP-2 (5 µg/ml) abolished the inhibitory effect of TIMP-2 (5 µg/ml) on MMP-2 (1 µg/ml) activity (assessed by [14C]-gelatin degradation). Overall, these results suggested that t-buOOH inactivated TIMP-2, the ambient inhibitor of MMP-2, leading to activation of the ambient proteinase, MMP-2 which subsequently stimulated Ca²⁺-ATPase activity and ATP-dependent Ca²⁺ uptake, but inhibited Na⁺-dependent Ca²⁺ uptake, resulting in a marked decrease in Ca²⁺ uptake in the microsomes. |
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Date |
2009-03-30T06:54:26Z
2009-03-30T06:54:26Z 2005-02 |
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Type |
Article
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Identifier |
0301-1208
http://hdl.handle.net/123456789/3502 |
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Language |
en_US
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Relation |
C12N 9/00
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Publisher |
CSIR
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Source |
IJBB Vol.42(1) [February 2005]
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