Changes in calcium-dependent protein kinase activity during in vitro tuberization in potato

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A soluble Ca2+-dependent protein kinase (CDPK) was purified to homogeneity in potato (Solanum tuberosum L.) plants. Potato CDPK was strictly dependent on Ca2+ (one-half maximal activation 0.6 μM) and phosphorylated a wide diversity of substrates, in which Syntide 2 was the best phosphate acceptor (M...

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Detalles Bibliográficos
Autores principales:	MacIntosh, G.C., Ulloa, R.M., Raíces, M., Téllez-Iñón, M.T.
Formato:	Artículo publishedVersion
Lenguaje:	Inglés
Publicado:	1996
Materias:	Glycine max Solanum tuberosum Tuberosum
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_00320889_v112_n4_p1541_MacIntosh
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paperaa:paper_00320889_v112_n4_p1541_MacIntosh
record_format	dspace
spelling	paperaa:paper_00320889_v112_n4_p1541_MacIntosh2023-06-12T16:45:26Z Changes in calcium-dependent protein kinase activity during in vitro tuberization in potato PLANT PHYSIOL. 1996;112(4):1541-1550 MacIntosh, G.C. Ulloa, R.M. Raíces, M. Téllez-Iñón, M.T. Glycine max Solanum tuberosum Tuberosum A soluble Ca2+-dependent protein kinase (CDPK) was purified to homogeneity in potato (Solanum tuberosum L.) plants. Potato CDPK was strictly dependent on Ca2+ (one-half maximal activation 0.6 μM) and phosphorylated a wide diversity of substrates, in which Syntide 2 was the best phosphate acceptor (Michaelis constant = 30 μM). The kinase was inhibited by Ca2+- chelating agents, phenotiazine derivatives, and N-(6-aminohexyl)-5-chloro-1- naphthalenesulfonamide (one-half maximal inhibition = 0.25 mM). Polyclonal antibodies directed against the regulatory region of the soybean CDPK recognized a 53-kD polypeptide. In an autophosphorylation assay, this same band was strongly labeled with [γ-32P]ATP in the presence of Ca2+. CDPK activity was high in nontuberized plants, but increased 2.5-fold at the onset of tuber development and was reduced to one-half of its original activity when the tuber had completed formation. In the early stages of tuberization, Ca2+-dependent phosphorylation of endogenous targets (specific bands of 68, 51, and 46 kD) was observed. These polypeptides were not labeled in nontuberizing plants or in completely formed tubers, indicating that this phosphorylation is a stage-specific event. In addition, dephosphorylation of specific polypeptides was detected in tuberizing plants, suggesting the involvement of a phosphatase. Preincubation of crude extracts with phosphatase inhibitors rendered a 100% increase in CDPK activity. Fil:Raíces, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 1996 info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion application/pdf eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00320889_v112_n4_p1541_MacIntosh
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
language	Inglés
orig_language_str_mv	eng
topic	Glycine max Solanum tuberosum Tuberosum
spellingShingle	Glycine max Solanum tuberosum Tuberosum MacIntosh, G.C. Ulloa, R.M. Raíces, M. Téllez-Iñón, M.T. Changes in calcium-dependent protein kinase activity during in vitro tuberization in potato
topic_facet	Glycine max Solanum tuberosum Tuberosum
description	A soluble Ca2+-dependent protein kinase (CDPK) was purified to homogeneity in potato (Solanum tuberosum L.) plants. Potato CDPK was strictly dependent on Ca2+ (one-half maximal activation 0.6 μM) and phosphorylated a wide diversity of substrates, in which Syntide 2 was the best phosphate acceptor (Michaelis constant = 30 μM). The kinase was inhibited by Ca2+- chelating agents, phenotiazine derivatives, and N-(6-aminohexyl)-5-chloro-1- naphthalenesulfonamide (one-half maximal inhibition = 0.25 mM). Polyclonal antibodies directed against the regulatory region of the soybean CDPK recognized a 53-kD polypeptide. In an autophosphorylation assay, this same band was strongly labeled with [γ-32P]ATP in the presence of Ca2+. CDPK activity was high in nontuberized plants, but increased 2.5-fold at the onset of tuber development and was reduced to one-half of its original activity when the tuber had completed formation. In the early stages of tuberization, Ca2+-dependent phosphorylation of endogenous targets (specific bands of 68, 51, and 46 kD) was observed. These polypeptides were not labeled in nontuberizing plants or in completely formed tubers, indicating that this phosphorylation is a stage-specific event. In addition, dephosphorylation of specific polypeptides was detected in tuberizing plants, suggesting the involvement of a phosphatase. Preincubation of crude extracts with phosphatase inhibitors rendered a 100% increase in CDPK activity.
format	Artículo Artículo publishedVersion
author	MacIntosh, G.C. Ulloa, R.M. Raíces, M. Téllez-Iñón, M.T.
author_facet	MacIntosh, G.C. Ulloa, R.M. Raíces, M. Téllez-Iñón, M.T.
author_sort	MacIntosh, G.C.
title	Changes in calcium-dependent protein kinase activity during in vitro tuberization in potato
title_short	Changes in calcium-dependent protein kinase activity during in vitro tuberization in potato
title_full	Changes in calcium-dependent protein kinase activity during in vitro tuberization in potato
title_fullStr	Changes in calcium-dependent protein kinase activity during in vitro tuberization in potato
title_full_unstemmed	Changes in calcium-dependent protein kinase activity during in vitro tuberization in potato
title_sort	changes in calcium-dependent protein kinase activity during in vitro tuberization in potato
publishDate	1996
url	http://hdl.handle.net/20.500.12110/paper_00320889_v112_n4_p1541_MacIntosh
work_keys_str_mv	AT macintoshgc changesincalciumdependentproteinkinaseactivityduringinvitrotuberizationinpotato AT ulloarm changesincalciumdependentproteinkinaseactivityduringinvitrotuberizationinpotato AT raicesm changesincalciumdependentproteinkinaseactivityduringinvitrotuberizationinpotato AT tellezinonmt changesincalciumdependentproteinkinaseactivityduringinvitrotuberizationinpotato
_version_	1769810224110108672

Changes in calcium-dependent protein kinase activity during in vitro tuberization in potato

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