INVESTIGADORES
ULLOA DE LA SERNA Rita Maria
artículos
Título:
Changes in Calcium dependent protein kinase activity during in vitro tuberization in potato
Autor/es:
MACINTOSH G; ULLOA RM; RAICES M; TÉLLEZ-IÑÓN MT
Revista:
PLANT PHYSIOLOGY.
Editorial:
ASPB
Referencias:
Año: 1996 vol. 112 p. 1541 - 1550
ISSN:
0032-0889
Resumen:
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(Solanum tuberosum L.) plants. Potato CDPK
was strictly dependent on Ca2+ (one-half maximal activation 0.6
p ~an)d phosphorylated a wide diversity of substrates, in which
Syntide 2 was the best phosphate acceptor (Michaelis constant = 30
p ~ )T.h e kinase was inhibited by Ca2+-chelating agents, phenotiazine
derivatives, and N-(6-aminohexyl)-5-chloro-l -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
[y3P1ATP 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 ofof
tuberization, Ca2+-dependent phosphorylation of endogenous targetsof 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.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.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.100% increase in CDPK activity.T.h e kinase was inhibited by Ca2+-chelating agents, phenotiazine
derivatives, and N-(6-aminohexyl)-5-chloro-l -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
[y3P1ATP 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 ofof
tuberization, Ca2+-dependent phosphorylation of endogenous targetsof 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.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.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.100% increase in CDPK activity.N-(6-aminohexyl)-5-chloro-l -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
[y3P1ATP 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 ofof
tuberization, Ca2+-dependent phosphorylation of endogenous targetsof 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.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.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.100% increase in CDPK activity.an)d phosphorylated a wide diversity of substrates, in which
Syntide 2 was the best phosphate acceptor (Michaelis constant = 30
p ~ )T.h e kinase was inhibited by Ca2+-chelating agents, phenotiazine
derivatives, and N-(6-aminohexyl)-5-chloro-l -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
[y3P1ATP 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 ofof
tuberization, Ca2+-dependent phosphorylation of endogenous targetsof 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.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.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.100% increase in CDPK activity.T.h e kinase was inhibited by Ca2+-chelating agents, phenotiazine
derivatives, and N-(6-aminohexyl)-5-chloro-l -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
[y3P1ATP 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 ofof
tuberization, Ca2+-dependent phosphorylation of endogenous targetsof 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.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.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.100% increase in CDPK activity.N-(6-aminohexyl)-5-chloro-l -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
[y3P1ATP 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 ofof
tuberization, Ca2+-dependent phosphorylation of endogenous targetsof 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.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.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.100% increase in CDPK activity.2 was the best phosphate acceptor (Michaelis constant = 30
p ~ )T.h e kinase was inhibited by Ca2+-chelating agents, phenotiazine
derivatives, and N-(6-aminohexyl)-5-chloro-l -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
[y3P1ATP 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 ofof
tuberization, Ca2+-dependent phosphorylation of endogenous targetsof 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.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.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.100% increase in CDPK activity.T.h e kinase was inhibited by Ca2+-chelating agents, phenotiazine
derivatives, and N-(6-aminohexyl)-5-chloro-l -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
[y3P1ATP 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 ofof
tuberization, Ca2+-dependent phosphorylation of endogenous targetsof 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.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.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.100% increase in CDPK activity.N-(6-aminohexyl)-5-chloro-l -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
[y3P1ATP 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 ofof
tuberization, Ca2+-dependent phosphorylation of endogenous targetsof 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.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.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.100% increase in CDPK activity.= 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
[y3P1ATP 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 ofof
tuberization, Ca2+-dependent phosphorylation of endogenous targetsof 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.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.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.100% increase in CDPK activity.