Molecular cloning, localization and characterization of a 40-kDa catecholamine-regulated protein.
We have previously described catecholamine-regulated proteins of molecular masses 47, 40 and 26 kDa (CRP47/40/26). In mammals, these proteins are detected only in brain and have been implicated as playing a role in dopaminergic neurotransmission. In this report, we have cloned the cDNA encoding CRP40 from bovine brain. Analysis of the predicted amino acid sequence revealed that the CRP40 product contains an hsp70 motif and shares homology with heat-shock protein hsp70. Immunolocalization studies using mAbs to dopamine show that it colocalizes with CRP40 in the vesicles of dopaminergic neuroblastoma SH-SY5Y cells. The constitutive expression of CRP40 was increased by exposure to heat shock similar to inducible heat-shock protein hsp70 in SH-SY5Y cells. Dopamine significantly modulated the levels of CRP40, whereas, the expression of hsp70 remained unchanged upon dopamine treatment of these cells. Moreover, CRP40 is able to prevent the thermal aggregation of luciferase in vitro, similar to hsp70, suggesting that CRP40 encodes a dopamine-inducible protein with properties similar to heat-shock proteins. The immunofluorescence analyses show that in SH-SY5Y cells, CRP40 translocates to the nucleus during dopamine-induced apoptosis. These results suggest that CRP40 could play a protective role against the harmful effects of catecholamine metabolites.
Nair
V D
VD
Departments of Psychiatry and Behavioral Neurosciences, McMaster University, Hamilton, Ontario, Canada. venugn01@doc.mssm.edu
Mishra
R K
RK
D000595:Amino Acid Sequence / Q000235:genetics; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis; D002395:Catecholamines / Q000378:metabolism* / Q000494:pharmacology; D002417:Cattle; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003001:Cloning, Molecular; D004298:Dopamine / Q000378:metabolism / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D015870:Gene Expression / Q000187:drug effects; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D017434:Protein Structure, Tertiary / Q000235:genetics; D021381:Protein Transport; D051381:Rats; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014162:Transfection
D000975:Antioxidants; D002395:Catecholamines; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D009419:Nerve Tissue Proteins; C422207:catecholamine-regulated protein, 40 kDa; D008156:Luciferases; D004298:Dopamine