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Abstract:

cAMP sites of the cAMP‐dependent protein kinase from the fungus Mucor rouxii have been characterized through the study of the effects of cAMP and of cAMP analogs on the phosphotransferase activity and through binding kinetics. The tetrameric holoenzyme, which contains two regulatory (R) and two catalytic (C) subunits, exhibited positive cooperativity in activation by cAMP, suggesting multiple cAMP‐binding sites. Several other results indicated that the Mucor kinase contained two different cooperative cAMP‐binding sites on each R subunit, with properties similar to those of the mammalian cAMP‐dependent protein kinase. Under optimum binding conditions, the [3H]cAMP dissociation behavior indicated equal amounts of two components which had dissociation rate constants of 0.09 min−1 (site 1) and 0.90 min−1 (site 2) at 30°C. Two cAMP‐binding sites could also be distinguished by C‐8 cAMP analogs (site‐1‐selective) and C‐6 cAMP analogs (site‐2‐selective); combinations of site‐1‐ and site‐2‐selective analogs were synergistic in protein kinase activation. The two different cooperative binding sites were probably located on the same R subunit, since the proteolytically derived dimeric form of the enzyme, which contained one R and one C component, retained the salient properties of the untreated tetrameric enzyme. Unlike any of the mammalian cyclic‐nucleotide‐dependent isozymes described thus far, the Mucor kinase was much more potently activated by C‐6 cAMP analogs than by C‐8 cAMP analogs. In the ternary complex formed by the native Mucor tetramer and cAMP, only the two sites 1 contained bound cAMP, a feature which has also not yet been demonstrated for the mammalian cAMP‐dependent protein kinase. Copyright © 1989, Wiley Blackwell. All rights reserved

Registro:

Documento: Artículo
Título:Two different intrachain cAMP sites in the cAMP‐dependent protein kinase of the dimorphic fungus Mucor rouxii
Autor:Paveto, C.; Passeron, S.; Corbin, J.D.; Moreno, S.
Filiación:Departamento de Quimica Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Argentina
Howard Hughes Medical Institute Laboratory, Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, United States
Palabras clave:cyclic amp; cyclic amp dependent protein kinase; cyclic amp derivative; radioisotope; binding site; fungus; mucor rouxii; nonhuman; priority journal; Allosteric Regulation; Binding Sites; Catalysis; Centrifugation, Density Gradient; Cyclic AMP; Enzyme Activation; Hydrolysis; Isoenzymes; Mucor; Protein Binding; Protein Kinases; Support, Non-U.S. Gov't; Trypsin
Año:1989
Volumen:179
Número:2
Página de inicio:429
Página de fin:434
DOI: http://dx.doi.org/10.1111/j.1432-1033.1989.tb14571.x
Título revista:European Journal of Biochemistry
Título revista abreviado:Eur. J. Biochem.
ISSN:00142956
CAS:cyclic AMP, 60-92-4; Cyclic AMP, 60-92-4; Isoenzymes; Protein Kinases, EC 2.7.1.37; Trypsin, EC 3.4.21.4
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00142956_v179_n2_p429_Paveto

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Citas:

---------- APA ----------
Paveto, C., Passeron, S., Corbin, J.D. & Moreno, S. (1989) . Two different intrachain cAMP sites in the cAMP‐dependent protein kinase of the dimorphic fungus Mucor rouxii. European Journal of Biochemistry, 179(2), 429-434.
http://dx.doi.org/10.1111/j.1432-1033.1989.tb14571.x
---------- CHICAGO ----------
Paveto, C., Passeron, S., Corbin, J.D., Moreno, S. "Two different intrachain cAMP sites in the cAMP‐dependent protein kinase of the dimorphic fungus Mucor rouxii" . European Journal of Biochemistry 179, no. 2 (1989) : 429-434.
http://dx.doi.org/10.1111/j.1432-1033.1989.tb14571.x
---------- MLA ----------
Paveto, C., Passeron, S., Corbin, J.D., Moreno, S. "Two different intrachain cAMP sites in the cAMP‐dependent protein kinase of the dimorphic fungus Mucor rouxii" . European Journal of Biochemistry, vol. 179, no. 2, 1989, pp. 429-434.
http://dx.doi.org/10.1111/j.1432-1033.1989.tb14571.x
---------- VANCOUVER ----------
Paveto, C., Passeron, S., Corbin, J.D., Moreno, S. Two different intrachain cAMP sites in the cAMP‐dependent protein kinase of the dimorphic fungus Mucor rouxii. Eur. J. Biochem. 1989;179(2):429-434.
http://dx.doi.org/10.1111/j.1432-1033.1989.tb14571.x