The calcium-binding activity of a vacuole-associated, dehydrin-like protein is regulated by phosphorylation.
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.
Heyen
Bruce J
BJ
Department of Biology, Indiana University-Purdue University at Indianapolis, 723 West Michigan Street, Indianapolis, IN 46202-5132, USA.
Alsheikh
Muath K
MK
Smith
Elizabeth A
EA
Torvik
Carl F
CF
Seals
Darren F
DF
Randall
Stephen K
SK
D000135:Acid Phosphatase / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000378:metabolism; D000595:Amino Acid Sequence; D000077102:Apium / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D002118:Calcium / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D010766:Phosphorylation / Q000187:drug effects; D010770:Phosphotransferases / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D014617:Vacuoles / Q000378:metabolism*
D002135:Calcium-Binding Proteins; D010940:Plant Proteins; C075590:dehydrin-like protein, plant; D010770:Phosphotransferases; D000469:Alkaline Phosphatase; D000135:Acid Phosphatase; D002118:Calcium