Soluble guanylate cyclase purified from bovine lung contains heme and copper

Gerzer, R.; Böhme, Eycke; Hofmann, Franz; Schultz, Günter

doi:10.1016/0014-5793(81)80429-2

Cited by 312 publications

(141 citation statements)

References 11 publications

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“…We found 0.9 equivalents of heme per sGC heterodimer. These observations confirm earlier findings (7,19,33) and demonstrate that the recombinant human sGC purified in these studies is a fully functional sGC.…”

Section: Resultssupporting

confidence: 92%

“…sGC is a heterodimer, composed of 73-to 82-kDa ␣-and 70-kDa ␤-subunits (5, 6), which contains one heme prosthetic group per heterodimer (7). At least two isoforms for each subunit have been cloned from mammals (8)(9)(10)(11)(12), and sGC homologues were identified in other species, including medaka fish (13), insects Drosophila melanogaster (14), Anopheles sp.…”

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confidence: 99%

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Human recombinant soluble guanylyl cyclase: Expression, purification, and regulation

Lee¹,

Martin²,

Murad³

2000

Proc. Natl. Acad. Sci. U.S.A.

128

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The ␣1-and ␤1-subunits of human soluble guanylate cyclase (sGC) were coexpressed in the Sf9 cells͞baculovirus system. In addition to the native enzyme, constructs with hexahistidine tag at the amino and carboxyl termini of each subunit were coexpressed. This permitted the rapid and efficient purification of active recombinant enzyme on a nickel-affinity column. The enzyme has one heme per heterodimer and was readily activated with the NO donor sodium nitroprusside or 3- (

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Section: Resultssupporting

confidence: 92%

mentioning

confidence: 99%

Human recombinant soluble guanylyl cyclase: Expression, purification, and regulation

Lee¹,

Martin²,

Murad³

2000

Proc. Natl. Acad. Sci. U.S.A.

128

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“…The effect of BCS on the activity of S-nitrosothiols cannot therefore be explained by interference with redox regulation of soluble guanylate cyclase (White et al, 1976) or cyclic GMP-dependent protein kinase (Landgraf et al, 1991), nor by binding of copper in the guanylate cyclase enzyme (Gerzer et al, 1981). In addition, the likelihood that BCS is acting by some non-specific mechanism is weakened by its failure to influence platelet inhibition by prostacyclin.…”

Section: Discussionmentioning

confidence: 99%

Copper chelation‐induced reduction of the biological activity of S‐nitrosothiols

Gordge

Meyer

Hothersall

et al. 1995

British J Pharmacology

100

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1 The effect of copper on the activity of the S-nitrosothiol compounds S-nitrosocysteine (cysNO) and S-nitrosoglutathione (GSNO) was investigated, using the specific copper chelator bathocuproine sulphonate (BCS), and human washed platelets as target cells.2 Chelation of trace copper with BCS (10 gLM) in washed platelet suspensions reduced the inhibition of thrombin-induced platelet aggregation by GSNO; however, BCS had no significant effect on the anti-aggregatory action of cysNO. BCS inhibited cyclic GMP generation in response to both cysNO and GSNO. 3 The effect of BCS was rapid (within 30 s), and could be abolished by increasing the platelet concentration to 500 x 109 1 -'. 4 In BCS-treated platelet suspensions, the addition of Cu2" ions (0.37-2.37 fiM) led to a restoration of both guanylate cyclase activation and platelet aggregation inhibition by GSNO. 5 The anti-aggregatory activity of GSNO was reduced in a concentration-dependent manner by the copper (I)-specific chelators BCS and neocuproine, and to a smaller extent by desferal.No effect was observed with the copper (II) specific chelator, cuprizone, the iron-specific chelator, bathophenanthroline sulphonate, or the broader-specificity copper chelator, D-penicillamine. 6 In both BCS-treated and -untreated platelet suspensions, cys NO was more potent than GSNO as a stimulator of guanylate cyclase. In BCS-treated platelet suspensions there was no significant difference between the anti-aggregatory potency of cysNO and GSNO; however, in untreated suspensions, GSNO was significantly more potent than cysNO. Thus, when copper was available, GSNO produced a greater inhibition of aggregation than cysNO, despite being a less potent activator of guanylate cyclase. 7 The breakdown of cysNO and GSNO was measured spectrophotometrically by decrease in absorbance at 334 nm. In Tyrode buffer, cysNO (10 !LM) broke down at a rate of 3.3 tAM min-'. BCS (10 tLM) reduced this to 0.5 ZlM min-'. GSNO, however, was stable, showing no fall in absorbance over a period of 7 min even in the absence of BCS. 8 We conclude that copper is required for the activity of both cysNO and GSNO, although its influence on anti-aggregatory activity is only evident with GSNO. The stimulatory effect of copper is unlikely to be explained solely by catalysis of S-nitrosothiol breakdown. The enhancement by copper of th anti-aggregatory activity of GSNO, relative to cysNO, suggests that copper may be required for biological activity of GSNO which is independent of guanylate cyclase stimulation.

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“…Current views of sGC structure define a hemecontaining regulatory domain, formed by N-terminal portions of both subunits, and a catalytic domain formed by the C-terminal half of both subunits (23). It is postulated that NO binding induces changes in the regulatory domain (24,25) that are transmitted through a yet-unknown mechanism to the catalytic site, which results in a significant increase in V max of the enzyme and some decrease in the K m for GTP substrate (15,26,27).…”

mentioning

confidence: 99%

A constitutively activated mutant of human soluble guanylyl cyclase (sGC): Implication for the mechanism of sGC activation

Martin¹,

Sharina²,

Kots³

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

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Soluble guanylate cyclase purified from bovine lung contains heme and copper

Cited by 312 publications

References 11 publications

Human recombinant soluble guanylyl cyclase: Expression, purification, and regulation

Human recombinant soluble guanylyl cyclase: Expression, purification, and regulation

Copper chelation‐induced reduction of the biological activity of S‐nitrosothiols

A constitutively activated mutant of human soluble guanylyl cyclase (sGC): Implication for the mechanism of sGC activation

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