The 32 kDa tonoplast polypeptide Di associated with the V0‐type H+‐ATPase of Mesembryanthemum crystallinum L. in the CAM state: A proteolytically processed subunit B?

Zhigang, An; Löw, Rainer; Rausch, Thomas; Lüttge, Ulrich; Ratajczak, Rafael

doi:10.1016/0014-5793(96)00556-x

Cited by 33 publications

(12 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In the case of subunit VHA-B, two spots were identified (spots 318 and 414) that differed in molecular mass (32 and 26 kD, respectively). While the deduced molecular mass of VHA-B is 55 kD, it is known that this subunit can under certain conditions undergo in vivo proteolytic processing into two subunits with apparent molecular masses of 31 to 32 kD and 27 to 28 kD (Zhigang et al, 1996;Krisch et al, 2000;Ratajczak, 2000), matching the size of the proteins identified in this study. This phenomenon is not limited to M. crystallinum, as the V-ATPase B subunit of yeast, Vma2p, was also susceptible to proteolytic processing with the appearance of a 30-kD protein (Landolt-Marticorena et al, 1999).…”

Section: Identification Of Differentially Regulated Tonoplast Proteinsupporting

confidence: 69%

Quantitative Proteomics of the Tonoplast Reveals a Role for Glycolytic Enzymes in Salt Tolerance

et al. 2009

View full text Add to dashboard Cite

To examine the role of the tonoplast in plant salt tolerance and identify proteins involved in the regulation of transporters for vacuolar Na + sequestration, we exploited a targeted quantitative proteomics approach. Two-dimensional differential in-gel electrophoresis analysis of free flow zonal electrophoresis separated tonoplast fractions from control, and salt-treated Mesembryanthemum crystallinum plants revealed the membrane association of glycolytic enzymes aldolase and enolase, along with subunits of the vacuolar H + -ATPase V-ATPase. Protein blot analysis confirmed coordinated salt regulation of these proteins, and chaotrope treatment indicated a strong tonoplast association. Reciprocal coimmunoprecipitation studies revealed that the glycolytic enzymes interacted with the V-ATPase subunit B VHA-B, and aldolase was shown to stimulate V-ATPase activity in vitro by increasing the affinity for ATP. To investigate a physiological role for this association, the Arabidopsis thaliana cytoplasmic enolase mutant, los2, was characterized. These plants were salt sensitive, and there was a specific reduction in enolase abundance in the tonoplast from salt-treated plants. Moreover, tonoplast isolated from mutant plants showed an impaired ability for aldolase stimulation of V-ATPase hydrolytic activity. The association of glycolytic proteins with the tonoplast may not only channel ATP to the V-ATPase, but also directly upregulate H + -pump activity.

show abstract

Section: Identification Of Differentially Regulated Tonoplast Proteinsupporting

confidence: 69%

Quantitative Proteomics of the Tonoplast Reveals a Role for Glycolytic Enzymes in Salt Tolerance

et al. 2009

View full text Add to dashboard Cite

show abstract

“…6). The isolation of the PM, ER, and tonoplast fractions was confirmed by probing them with anti-H 1 -ATPase antibodies (Pardo and Serrano, 1989), anti-BiP antibodies (Anderson et al, 1994a(Anderson et al, , 1994b, and anti-V-ATPase antibodies (Zhigang et al, 1996), respectively. The isolation of the nuclear fraction was confirmed by probing it with anti-Ras-related GTPase (RAN) antibodies (Melchior et al, 1993;Moore and Blobel, 1993;Melchior and Gerace, 1998).…”

Section: Subcellular Fractionationmentioning

confidence: 99%

“…Anti-H 1 -ATPase antibody was used to identify the PM fraction (1:5,000 dilution in TBS, kindly provided by Dr. Serrano, Camino de Vera, Valencia, Spain; Pardo and Serrano, 1989). Anti-V-ATPase subunit A antibody was used to identify the tonoplast fraction (1:500 dilution in TBS, kindly provided by Dr. Ratajczak, Technische Hochschule Darmstadt, Darmstadt, Germany; Zhigang et al, 1996). Anti-NPC antibody was used to target nuclei (1:200 dilution in TBS; BabCO, Richmond, CA).…”

Section: Western-blot Analysismentioning

confidence: 99%

Autophosphorylation and Subcellular Localization Dynamics of a Salt- and Water Deficit-Induced Calcium-Dependent Protein Kinase from Ice Plant

et al. 2004

View full text Add to dashboard Cite

A salinity and dehydration stress-responsive calcium-dependent protein kinase (CDPK) was isolated from the common ice plant (Mesembryanthemum crystallinum; McCPK1). McCPK1 undergoes myristoylation, but not palmitoylation in vitro. Removal of the N-terminal myristate acceptor site partially reduced McCPK1 plasma membrane (PM) localization as determined by transient expression of green fluorescent protein fusions in microprojectile-bombarded cells. Removal of the N-terminal domain (amino acids 1-70) completely abolished PM localization, suggesting that myristoylation and possibly the N-terminal domain contribute to membrane association of the kinase. The recombinant, Escherichia coli-expressed, full-length McCPK1 protein was catalytically active in a calcium-dependent manner (K 0.5 5 0.15 mM). Autophosphorylation of recombinant McCPK1 was observed in vitro on at least two different Ser residues, with the location of two sites being mapped to Ser-62 and Ser-420. An Ala substitution at the Ser-62 or Ser-420 autophosphorylation site resulted in a slight increase in kinase activity relative to wild-type McCPK1 against a histone H1 substrate. In contrast, Ala substitutions at both sites resulted in a dramatic decrease in kinase activity relative to wild-type McCPK1 using histone H1 as substrate. McCPK1 undergoes a reversible change in subcellular localization from the PM to the nucleus, endoplasmic reticulum, and actin microfilaments of the cytoskeleton in response to reductions in humidity, as determined by transient expression of McCPK1-green fluorescent protein fusions in microprojectile-bombarded cells and confirmed by subcellular fractionation and western-blot analysis of 63 His-tagged McCPK1.Calcium is a ubiquitous and pivotal second messenger in the signal transduction networks that plants use to respond to a wide variety of physiological stimuli. Cytosolic Ca 21 fluctuations have been observed in response to a number of stimuli, including red light, abscisic acid (ABA), GA, drought, hyperand hypo-osmotic stress, ionic stress, touch, cold, heat shock, oxidative stress, fungal elicitors, and nodulation factors (Sanders et al., 2002 (Harmon et al., , 2001Hrabak, 2000;Hrabak et al., 2003).CDPKs are composed of a single polypeptide chain with a catalytic kinase domain at the N terminus, an intervening junction domain, and a calmodulin-like domain at the C terminus containing up to four functional Ca 21 -binding EF hands. The junction domain acts as an autoinhibitor in a pseudosubstrate fashion (Harper et al., 1994;Vitart et al., 2000;Weljie et al., 2000). Binding of Ca 21 to the calmodulin-like domain results in a conformational change leading to the release of the autoinhibitor domain from the active site and kinase activation (Harmon et al., 1994;Harper et al., 1994). Many CDPKs also have additional Nterminal leader and C-terminal domains composed of highly variable amino acid sequences. AtCPK25 and some CDPK-related kinases possess degenerate calmodulin domains (Lindzen and Choi, 1995;Zhang and Lu, 2003) and appa...

show abstract

“…Western blot analysis with an antiserum directed against the V-ATPase holoenzyme of KalanchoO" daigremontiana revealed a significant increase of the V-ATPase subunits A and B. Furthermore, levels of cross-reacting polypeptides in the range of 33-36 kDa were strongly elevated, which could either represent isoforms of subunit D [16] or stress-induced proteolytic degradation products of subunits A and/or B [ 1,2].…”

mentioning

confidence: 99%

Salt stress induces an increased expression of V-type H+-ATPase in mature sugar beet leaves

Kirsch¹,

Zhigang²,

Viereck³

et al. 1996

Plant Mol Biol

Self Cite

View full text Add to dashboard Cite

In the halotolerant sugar beet co-expression of V-ATPase and a vacuolar Na+/H(+)-antiporter provides a mechanism for vacuolar salt sequestration. To analyze salt-induced changes in the expression of the vacuolar H(+)-ATPase (V-ATPase) a partial cDNA of the proton-channel forming subunit c was cloned by RT-PCR. Southern blot analysis indicated a small gene family. In control plants transcript levels were high in roots and young growing leaves but low in fully expanded leaves. In mature leaves salt exposure (400 mM, 48 h) induced a strong increase in subunit c-mRNA. Transcripts for the catalytic subunit A followed a similar developmental and stress-modulated pattern, indicating a coordinate regulation of transcripts for both V-ATPase subunits. Concomittant with the mRNA increases the amount of V-ATPase protein increased as well.

show abstract

The 32 kDa tonoplast polypeptide D_i associated with the V0‐type H⁺‐ATPase of Mesembryanthemum crystallinum L. in the CAM state: A proteolytically processed subunit B?

Cited by 33 publications

References 17 publications

Quantitative Proteomics of the Tonoplast Reveals a Role for Glycolytic Enzymes in Salt Tolerance

Quantitative Proteomics of the Tonoplast Reveals a Role for Glycolytic Enzymes in Salt Tolerance

Autophosphorylation and Subcellular Localization Dynamics of a Salt- and Water Deficit-Induced Calcium-Dependent Protein Kinase from Ice Plant

Salt stress induces an increased expression of V-type H+-ATPase in mature sugar beet leaves

Contact Info

Product

Resources

About