The disappearance of an hsc70 species in mung bean seed during germination: purification and characterization of the protein.
We have purified a 73 kDa protein from the cytosolic fraction of mung bean seeds. It comprises 0.5-1% of the total protein in seeds. This purified protein is a bona fide hsc70 on the basis of several lines of evidence. First, antibodies against bovine brain hsc70 cross-react with the purified 73 kDa protein. Second, the purified protein comigrates on two-dimensional gels with one of the heat-inducible hsc70s in mung bean seedlings. Third, similar to other hsc70 species, the purified 73 kDa protein has a high affinity for ATP. Finally, the hydrolysis of ATP by the purified protein can be stimulated by peptides; ATPase activity increases from 40 nmol/h to 165 nmol/h per mg of protein. The purified mung bean hsc70 autophosphorylates at a substoichiometric level. Moreover, the amount of this hsc70 species diminishes while new species of hsc70s appear after germination, suggesting that the expression of hsc70 in mung bean is subject to developmental regulation.
Wang
C
C
Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan.
Lin
B L
BL
D000251:Adenosine Triphosphatases / Q000302:isolation & purification* / Q000378:metabolism*; D000595:Amino Acid Sequence; D002352:Carrier Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D002848:Chromatography, DEAE-Cellulose; D002966:Clathrin / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D007887:Fabaceae / Q000378:metabolism*; D050883:HSC70 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000378:metabolism; D010766:Phosphorylation; D010946:Plants, Medicinal; D012639:Seeds / Q000502:physiology*; D013379:Substrate Specificity; D014461:Ultracentrifugation
D002352:Carrier Proteins; D002966:Clathrin; D050883:HSC70 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D010446:Peptide Fragments; D000251:Adenosine Triphosphatases