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PMID: 8425059 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The disappearance of an hsc70 species in mung bean seed during germination: purification and characterization of the protein.

Plant molecular biology ·Vol. 21 ·No. 2 ·1993-01-00 ·Pages 317-29

Wang C, Lin BL

Abstract

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.

MeSH Terms
Adenosine Triphosphatases/isolation & purification,metabolism Amino Acid Sequence Carrier Proteins/isolation & purification,metabolism Chromatography, DEAE-Cellulose Clathrin/metabolism Cytosol/metabolism Electrophoresis, Gel, Two-Dimensional Electrophoresis, Polyacrylamide Gel Fabaceae/metabolism HSC70 Heat-Shock Proteins HSP70 Heat-Shock Proteins Heat-Shock Proteins/isolation & purification,metabolism Kinetics Molecular Sequence Data Molecular Weight Peptide Fragments/metabolism Phosphorylation Plants, Medicinal Seeds/physiology Substrate Specificity Ultracentrifugation
Chemicals
Carrier Proteins Clathrin HSC70 Heat-Shock Proteins HSP70 Heat-Shock Proteins Heat-Shock Proteins Peptide Fragments Adenosine Triphosphatases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Wang C
Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan.
Lin B L
References (46)
46 references, click to expand
  1. An enzyme that removes clathrin coats: purification of an uncoating ATPase.
    J Cell Biol. 1984 Aug;99(2):723-33 PMID: 6146630
  2. Nucleotide sequence of the cDNA of a bovine 70 kilodalton heat shock cognate protein.
    Nucleic Acids Res. 1990 Sep 25;18(18):5569 PMID: 2216746
  3. Identity of the immunoglobulin heavy-chain-binding protein with the 78,000-dalton glucose-regulated protein and the role of posttranslational modifications in its binding function.
    Mol Cell Biol. 1988 Oct;8(10):4250-6 PMID: 3141786
  4. 70-kD heat shock-related protein is one of at least two distinct cytosolic factors stimulating protein import into mitochondria.
    J Cell Biol. 1988 Dec;107(6 Pt 1):2051-7 PMID: 3058716
  5. Constitutively expressed rat mRNA encoding a 70-kilodalton heat-shock-like protein.
    Mol Cell Biol. 1985 Dec;5(12):3476-83 PMID: 3939319
  6. Protein kinase C autophosphorylates by an intrapeptide reaction.
    J Biol Chem. 1987 Jul 25;262(21):10185-8 PMID: 3611058
  7. Molecular chaperones.
    Annu Rev Biochem. 1991;60:321-47 PMID: 1679318
  8. Phosphorylation of the major heat shock protein of Dictyostelium discoideum.
    Mol Cell Biol. 1982 May;2(5):484-9 PMID: 6810097
  9. Developmental-stage-specific expression of the hsp70 gene family during differentiation of the mammalian male germ line.
    Mol Cell Biol. 1987 May;7(5):1791-6 PMID: 3600644
  10. The heat-shock proteins.
    Annu Rev Genet. 1988;22:631-77 PMID: 2853609
  11. The E. coli dnaK gene product, the hsp70 homolog, can reactivate heat-inactivated RNA polymerase in an ATP hydrolysis-dependent manner.
    Cell. 1990 Sep 7;62(5):939-44 PMID: 2203539
  12. Sequences of two hsc 70 cDNAs from Lycopersicon esculentum.
    Plant Mol Biol. 1991 Mar;16(3):475-8 PMID: 1893113
  13. Heat shock, stress proteins, chaperones, and proteotoxicity.
    Cell. 1991 Jul 26;66(2):191-7 PMID: 1855252
  14. Posttranslational association of immunoglobulin heavy chain binding protein with nascent heavy chains in nonsecreting and secreting hybridomas.
    J Cell Biol. 1986 May;102(5):1558-66 PMID: 3084497
  15. Uncoating protein (hsc70) binds a conformationally labile domain of clathrin light chain LCa to stimulate ATP hydrolysis.
    Cell. 1990 Sep 7;62(5):875-87 PMID: 1975516
  16. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
    Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4 PMID: 388439
  17. Isolation and characterization of a soybean hsp70 gene.
    Plant Mol Biol. 1991 Apr;16(4):671-83 PMID: 1714321
  18. The ATPase core of a clathrin uncoating protein.
    J Biol Chem. 1987 Jan 15;262(2):746-51 PMID: 3027066
  19. Heat shock proteins are methylated in avian and mammalian cells.
    Proc Natl Acad Sci U S A. 1981 Jun;78(6):3531-5 PMID: 6943552
  20. Characterization of an immunoglobulin binding protein homolog in the maize floury-2 endosperm mutant.
    Plant Cell. 1991 May;3(5):483-96 PMID: 1840923
  21. The dnaK protein of Escherichia coli possesses an ATPase and autophosphorylating activity and is essential in an in vitro DNA replication system.
    Proc Natl Acad Sci U S A. 1983 Nov;80(21):6431-5 PMID: 6314326
  22. Cyclic AMP-modulated phosphorylation of intermediate filament proteins in cultured avian myogenic cells.
    Mol Cell Biol. 1982 Sep;2(9):1104-14 PMID: 6294504
  23. Methylations of 70,000-Da heat shock proteins in 3T3 cells: alterations by arsenite treatment, by different stages of growth and by virus transformation.
    Arch Biochem Biophys. 1992 Aug 15;297(1):169-75 PMID: 1322111
  24. Interaction of DnaK with ATP: binding, hydrolysis and Ca+2-stimulated autophosphorylation.
    Biochem Biophys Res Commun. 1990 Feb 14;166(3):1284-92 PMID: 2106314
  25. Uncoating of coated vesicles by yeast hsp70 proteins.
    J Biol Chem. 1991 Oct 15;266(29):19565-71 PMID: 1833403
  26. Interactions of liver Grp78 and Escherichia coli recombinant Grp78 with ATP: multiple species and disaggregation.
    Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2081-5 PMID: 1532251
  27. Heat shock protein hsp70 cognate gene expression in vegetative and reproductive organs of Lycopersicon esculentum.
    Proc Natl Acad Sci U S A. 1989 May;86(10):3674-8 PMID: 16594044
  28. Protein folding in the cell.
    Nature. 1992 Jan 2;355(6355):33-45 PMID: 1731198
  29. Control of protein exit from the endoplasmic reticulum.
    Annu Rev Cell Biol. 1989;5:1-23 PMID: 2688704
  30. ADP-ribosylation of the Mr 83,000 stress-inducible and glucose-regulated protein in avian and mammalian cells: modulation by heat shock and glucose starvation.
    Proc Natl Acad Sci U S A. 1983 Aug;80(15):4664-8 PMID: 6576354
  31. Interaction of Hsp 70 with newly synthesized proteins: implications for protein folding and assembly.
    Science. 1990 May 18;248(4957):850-4 PMID: 2188360
  32. Peptide-binding specificity of the molecular chaperone BiP.
    Nature. 1991 Oct 24;353(6346):726-30 PMID: 1834945
  33. A subfamily of stress proteins facilitates translocation of secretory and mitochondrial precursor polypeptides.
    Nature. 1988 Apr 28;332(6167):800-5 PMID: 3282178
  34. 70K heat shock related proteins stimulate protein translocation into microsomes.
    Nature. 1988 Apr 28;332(6167):805-10 PMID: 3282179
  35. Immunocytolocalization of extensin in developing soybean seed coats by immunogold-silver staining and by tissue printing on nitrocellulose paper.
    J Cell Biol. 1987 Dec;105(6 Pt 1):2581-8 PMID: 3693394
  36. Loss of BiP/GRP78 function blocks translocation of secretory proteins in yeast.
    J Cell Biol. 1990 Jun;110(6):1885-95 PMID: 2190988
  37. Characterization of an HSP70 Cognate Gene Family in Arabidopsis.
    Plant Physiol. 1988 Nov;88(3):731-40 PMID: 16666375
  38. A role for a 70-kilodalton heat shock protein in lysosomal degradation of intracellular proteins.
    Science. 1989 Oct 20;246(4928):382-5 PMID: 2799391
  39. Calcium-dependent autophosphorylation of the glucose-regulated protein, Grp78.
    Arch Biochem Biophys. 1991 Sep;289(2):256-61 PMID: 1910317
  40. Functional domains of the Escherichia coli dnaK heat shock protein as revealed by mutational analysis.
    J Biol Chem. 1989 Dec 15;264(35):21122-30 PMID: 2531744
  41. High resolution two-dimensional electrophoresis of proteins.
    J Biol Chem. 1975 May 25;250(10):4007-21 PMID: 236308
  42. Three-dimensional structure of the ATPase fragment of a 70K heat-shock cognate protein.
    Nature. 1990 Aug 16;346(6285):623-8 PMID: 2143562
  43. Peptide binding and release by proteins implicated as catalysts of protein assembly.
    Science. 1989 Jul 28;245(4916):385-90 PMID: 2756425
  44. The tobacco luminal binding protein is encoded by a multigene family.
    Plant Cell. 1991 Sep;3(9):1025-35 PMID: 1822990
  45. Expression of wild-type and mutant forms of influenza hemagglutinin: the role of folding in intracellular transport.
    Cell. 1986 Sep 12;46(6):939-50 PMID: 3757030
  46. The structure and expression of maize genes encoding the major heat shock protein, hsp70.
    EMBO J. 1986 Mar;5(3):451-8 PMID: 16453670
Article Info
Journal
Plant molecular biology
Abbr.
Plant Mol Biol
ISSN
0167-4412
DOI
10.1007/BF00019947
Published
1993-01-00
Pages
317-29
Language
English
Region
Netherlands
NLM ID
9106343
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00019947 View journal details
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