Molecular and biochemical characterization of a calcium/calmodulin-binding protein kinase from rice

Zhang, Lei; Liu, Bi‐Feng; Liang, Shuping; Jones, Russell L.; Lu, Ying‐Tang

doi:10.1042/bj20020780

Cited by 68 publications

(57 citation statements)

References 44 publications

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“…This may reflect differences among organisms with respect to the cellular levels of NADK substrates. The high affinity of Arabidopsis NADK for CaM (S 0.5 of 2.2 nM) is consistent with reports on other plant NADKs (Harmon et al, 1984;Lee et al, 1997;Delumeau et al, 2000) and other plant CaM-binding proteins (Snedden et al, 1996;Bouche et al, 2002;Zhang et al, 2002;Chandok et al, 2003). Similarly, the S 0.5 value for Ca 21 (0.8 mM) suggests stimuli that elevate Ca 21 from resting levels in vivo may elicit a CaM-modulated NADK response.…”

Section: Discussionsupporting

confidence: 74%

Cloning and Characterization of Two NAD Kinases from Arabidopsis. Identification of a Calmodulin Binding Isoform

et al. 2004

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NAD kinase (NADK; ATP:NAD 2#-phosphotransferase, EC 2.7.1.23), an enzyme found in both prokaryotes and eukaryotes, generates the important pyridine nucleotide NADP from substrates ATP and NAD. The role of NADKs in plants is poorly understood, and cDNAs encoding plant NADKs have not previously been described to our knowledge. We have cloned two cDNAs from Arabidopsis predicted to encode NADK isoforms, designated NADK1 and NADK2, respectively. Expressed as recombinant proteins in bacteria, both NADK1 and NADK2 were catalytically active, thereby confirming their identity as NADKs. Transcripts for both isoforms were detected in all tissues examined and throughout development. Although the predicted catalytic regions for NADK1 and NADK2 show sequence similarity to NADKs from other organisms, NADK2 possesses a large N-terminal extension that appears to be unique to plants. Using recombinant glutathione-S-transferase fusion proteins and calmodulin (CaM)-affinity chromatography, we delineated a Ca 21 -dependent CaM-binding domain to a 45-residue region within the N-terminal extension of NADK2. Although recombinant NADK2 was not responsive to CaM in vitro, immunoblot analysis suggests that native NADK2 is a CaM-binding protein. In Arabidopsis crude extracts, CaMdependent NADK activity was much greater than CaM-independent activity throughout development, particularly in young seedlings. A native CaM-dependent NADK was partially purified from Arabidopsis seedlings (K m NAD 5 0.20 mM, K m Mg 21 2ATP 5 0.17 mM). The enzyme was fully activated by conserved CaM (S 0.5 5 2.2 nM) in the presence of calcium but displayed differential responsiveness to eight CaM-like Arabidopsis proteins. Possible roles for NADKs in plants are discussed in light of our observations.

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Section: Discussionsupporting

confidence: 74%

Cloning and Characterization of Two NAD Kinases from Arabidopsis. Identification of a Calmodulin Binding Isoform

et al. 2004

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“…An acidic condition of pH 6.0 and a basic condition of pH 9.5 reduced the kinase activity to 34% and 60% respectively. The pH-oriented activity of NtCPK5 was similar to that observed for OsCBK in rice [12]. NtCPK5 can phosphorylate histone IIIs rapidly and reached about 60% of the maximal activity within 10 min in the presence of 0.1 mM CaCl 2 (Fig.…”

Section: Characterization Of the Kinetic Properties Of Ntcpk5supporting

confidence: 74%

“…To identify the kinase properties of NtCPK5, several constructs were made in plasmid pFastHTb following the protocol described previously [12,13]. Using pNtCPK5 as template, the cDNAs for the full open reading frame (NtCPK5) and two truncated forms P1 (residues 1-372) and P2 (residues 1-408) of NtCPK5 were amplified and purified.…”

Section: Construction Of Plasmids Ntcpk5 and Truncated Ntcpk5 P1 And P2mentioning

confidence: 99%

Cellular localization and biochemical characterization of a novel calcium-dependent protein kinase from tobacco

Wang

Zhang

et al. 2005

Cell Res

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By screening tobacco cDNA library with MCK1 as a probe, we isolated a cDNA clone NtCPK5 (accession number AY971376), which encodes a typical calcium-dependent protein kinase. Sequence analyses indicated that NtCPK5 is related to both CPKs and CRKs superfamilies and has all of the three conserved domains of CPKs. The biochemical activity of NtCPK5 was calcium-dependent. NtCPK5 had V max and K m of 526 nmol/min/mg and 210 µg/ml respectively with calf thymus histone (fraction III, abbreviated to histone IIIs) as substrate. For substrate syntide-2, NtCPK5 showed a higher V max of 2008 nmol/min/mg and a lower K m of 30 µM. The K 0.5 of calcium activation was 0.04 µM or 0.06 µM for histone IIIs or syntide-2 respectively. The putative myristoylation and palmitoylation consensus sequence of NtCPK5 suggests that it could be a membrane-anchoring protein. Indeed, our transient expression experiments with wild type and mutant forms of NtCPK5/GFP fusion proteins showed that NtCPK5 was localized to the plasma membrane of onion epidermal cells and that the localization required the N-terminal acylation sites of NtCPK5/GFP. Taking together, our data have demonstrated the biochemical characteristics of a novel protein NtCPK5 and its subcellular localization as a membrane-anchoring protein.

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“…In addition, it does not appear that CRKs contain functional autoinhibitory domains, because a maize CRK was equally active in the presence of calcium or EGTA (Furumoto et al, 1996). A rice CRK binds calmodulin in a calcium-dependent manner, although its enzymatic activity is independent of calcium and calmodulin (Zhang et al, 2002). Thus, CRKs may be constitutively active.…”

Section: Crkmentioning

confidence: 99%

The Arabidopsis CDPK-SnRK Superfamily of Protein Kinases

Hrabak

Chan²,

Gribskov³

et al. 2003

Plant Physiology

898

823

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The CDPK-SnRK superfamily consists of seven types of serine-threonine protein kinases: calcium-dependent protein kinase (CDPKs), CDPK-related kinases (CRKs), phosphoenolpyruvate carboxylase kinases (PPCKs), PEP carboxylase kinase-related kinases (PEPRKs), calmodulin-dependent protein kinases (CaMKs), calcium and calmodulin-dependent protein kinases (CCaMKs), and SnRKs. Within this superfamily, individual isoforms and subfamilies contain distinct regulatory domains, subcellular targeting information, and substrate specificities. Our analysis of the Arabidopsis genome identified 34 CDPKs, eight CRKs, two PPCKs, two PEPRKs, and 38 SnRKs. No definitive examples were found for a CCaMK similar to those previously identified in lily (Lilium longiflorum) and tobacco (Nicotiana tabacum) or for a CaMK similar to those in animals or yeast. CDPKs are present in plants and a specific subgroup of protists, but CRKs, PPCKs, PEPRKs, and two of the SnRK subgroups have been found only in plants. CDPKs and at least one SnRK have been implicated in decoding calcium signals in Arabidopsis. Analysis of intron placements supports the hypothesis that CDPKs, CRKs, PPCKs and PEPRKs have a common evolutionary origin; however there are no conserved intron positions between these kinases and the SnRK subgroup. CDPKs and SnRKs are found on all five Arabidopsis chromosomes. The presence of closely related kinases in regions of the genome known to have arisen by genome duplication indicates that these kinases probably arose by divergence from common ancestors. The PlantsP database provides a resource of continuously updated information on protein kinases from Arabidopsis and other plants.In eukaryotes, protein kinases are involved in regulating key aspects of cellular function, including cell division, metabolism, and responses to external signals. The completed sequence of the Arabidopsis genome provides the first opportunity to identify all of the protein kinases present in a model plant. The Arabidopsis genome encodes 1,085 typical protein kinases (M. Gribskov, unpublished data), which is about 4% of the predicted 25,500 genes (Arabidopsis Article, publication date, and citation information can be found at www.plantphysiol.org/cgi

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Molecular and biochemical characterization of a calcium/calmodulin-binding protein kinase from rice

Cited by 68 publications

References 44 publications

Cloning and Characterization of Two NAD Kinases from Arabidopsis. Identification of a Calmodulin Binding Isoform

Cloning and Characterization of Two NAD Kinases from Arabidopsis. Identification of a Calmodulin Binding Isoform

Cellular localization and biochemical characterization of a novel calcium-dependent protein kinase from tobacco

The Arabidopsis CDPK-SnRK Superfamily of Protein Kinases

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