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PMID: 12376635 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

The calcium-binding activity of a vacuole-associated, dehydrin-like protein is regulated by phosphorylation.

Plant physiology ·Vol. 130 ·No. 2 ·2002-10-00 ·Pages 675-87

Heyen BJ, Alsheikh MK, Smith EA, Torvik CF, Seals DF, Randall SK

Abstract

A vacuole membrane-associated calcium-binding protein with an apparent mass of 45 kD was purified from celery (Apium graveolens). This protein, VCaB45, is enriched in highly vacuolate tissues and is located within the lumen of vacuoles. Antigenically related proteins are present in many dicotyledonous plants. VCaB45 contains significant amino acid identity with the dehydrin family signature motif, is antigenically related to dehydrins, and has a variety of biochemical properties similar to dehydrins. VCaB45 migrates anomalously in sodium dodecyl sulfate-polyacrylamide gel electrophoresis having an apparent molecular mass of 45 kD. The true mass as determined by matrix-assisted laser-desorption ionization time of flight was 16.45 kD. VCaB45 has two characteristic dissociation constants for calcium of 0.22 +/- 0.142 mM and 0.64 +/- 0.08 mM, and has an estimated 24.7 +/- 11.7 calcium-binding sites per protein. The calcium-binding properties of VCaB45 are modulated by phosphorylation; the phosphorylated protein binds up to 100-fold more calcium than the dephosphorylated protein. VCaB45 is an "in vitro" substrate of casein kinase II (a ubiquitous eukaryotic kinase), the phosphorylation resulting in a partial activation of calcium-binding activity. The vacuole localization, calcium binding, and phosphorylation of VCaB45 suggest potential functions.

MeSH Terms
Acid Phosphatase/metabolism Alkaline Phosphatase/metabolism Amino Acid Sequence Apium/genetics,metabolism Blotting, Western Calcium/pharmacology Calcium-Binding Proteins/drug effects,genetics,metabolism Electrophoresis, Polyacrylamide Gel Molecular Sequence Data Phosphorylation/drug effects Phosphotransferases/metabolism Plant Proteins/genetics,metabolism Sequence Homology, Amino Acid Vacuoles/metabolism
Chemicals
Calcium-Binding Proteins Plant Proteins dehydrin-like protein, plant Phosphotransferases Alkaline Phosphatase Acid Phosphatase Calcium
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Heyen Bruce J
Department of Biology, Indiana University-Purdue University at Indianapolis, 723 West Michigan Street, Indianapolis, IN 46202-5132, USA.
Alsheikh Muath K
Smith Elizabeth A
Torvik Carl F
Seals Darren F
Randall Stephen K
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Article Info
Journal
Plant physiology
Abbr.
Plant Physiol
ISSN
0032-0889
DOI
10.1104/pp.002550
Published
2002-10-00
Pages
675-87
Language
English
Region
United States
NLM ID
0401224
PMCID
PMC166597
Subset
IM
External Links
PubMed source DOI: 10.1104/pp.002550 PMC Full Text View journal details
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