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p19 detected in the rat retina and pineal gland is a guanylyl cyclase-activating protein [GCAP]

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Dejda A., Matczak I., Gorczyca W. A.

Acta Biochimica Polonica

2002

tom 49

nr 4

The Ca2+ -dependent activation of retina-specific guanylyl cyclase (retGC) is mediated by guanylyl cyclase-activating proteins (GCAPs). Here we report for the first time detection of a 19 kDa protein (p19) with GCAP properties in extracts of rat retina and pineal gland. Both extracts stimulate synthesis of cGMP in rod outer segment (ROS) membranes at low (30 nM) but not at high (1 uM) concentrations of Ca2+ .AtlowCa2+ , immunoaffinity purified p19 activates guanylyl cyclase(s) in bovine ROS and rat retinal membranes. Moreover, p19 is recognized by antibodies against bovine GCAP1 and, similarly to other GCAPs, exhibits a Ca2+ -dependent electrophoretic mobility shift.

Ca2plus differently affects hydrophobic properties of guanylyl cyclase-activating proteins [GCAPs] and recoverin

84%

Gorczyca W. A., Kobialka M., Kuropatwa M., Kurowska E.

Acta Biochimica Polonica

2003

tom 50

nr 2

Guanylyl cyclase-activating proteins (GCAPs) and recoverin are retina-specific Ca2+ -binding proteins involved in phototransduction. We provide here evidence that in spite of structural similarities GCAPs and recoverin differently change their overall hydrophobic properties in response to Ca2+ . Using native bovine GCAP1, GCAP2 and recoverin we show that: i) the Ca2+ -dependent binding of recoverin to Phenyl-Se- pharose is distinct from such interactions of GCAPs; ii) fluorescence intensity of 1-anilinonaphthalene-8-sulfonate (ANS) is markedly higher at high [Ca2+ ]free (10 uM) than at low [Ca2+ ]freee(10 nM) in the presence of recoverin, while an opposing effect is observed in the presence of GCAPs; iii) fluorescence resonance energy transfer from tryptophane residues to ANS is more efficient at high [Ca2+ ]free in recoverin and at low [Ca2+ ]freee in GCAP2. Such different changes of hydrophobicity evoked by Ca2+ appear to be the precondition for possible mechanisms by which GCAPs and recoverin control the activities of their target enzymes.

Ograniczanie wyników

2 Acta Biochimica Polonica

2 Gorczyca W. A.

1 Dejda A.

1 Kobialka M.

1 Kuropatwa M.

1 Kurowska E.

1 Matczak I.

1 2003

1 2002

Wyniki wyszukiwania

p19 detected in the rat retina and pineal gland is a guanylyl cyclase-activating protein [GCAP]

Ca2plus differently affects hydrophobic properties of guanylyl cyclase-activating proteins [GCAPs] and recoverin