Calreticulin--a stress protein induced in the renal epithelial cell line NBL-1 by amino acid deprivation.
Confluent monolayer cultures of the bovine kidney cell line NBL-1 were starved of amino acids in the presence of tracer concentrations of [35S]-methionine. Fluorographs of SDS-polyacrylamide gel separated membrane proteins revealed increased labelling of at least two proteins in starved cells relative to those in cells grown in complete medium. The patterns of Coomassie blue stained proteins from Concanavalin A-purified fractions of cells grown under fed and amino acid-starved conditions were similar but fluorography indicated the presence of one major labelled glycoprotein with a molecular weight of 62 kD in starved cells which was not present in fed cells. N-terminal amino acid analysis of the first 15 amino acids of the 62 kD protein and a protein of 60 kD found in control cells identified both proteins as calreticulin. N-terminal amino acid sequence analysis of a second amino acid starvation-up-regulated protein identified it as glucose-regulated protein GRP78. The amino acid sequences of calreticulin, GRP78 and two transport proteins known to be induced in amino acid starvation, have a common motif near the C-terminal end of the molecule. It is suggested that calreticulin is a member of a novel class of stress proteins induced by amino acid starvation.
Department of Biochemistry, School of Medical Sciences, Bristol, UK.
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S
Plakidou-Dymock
J D
JD
McGivan
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000378:metabolism* / Q000494:pharmacology*; D000818:Animals; D015153:Blotting, Western; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D037282:Calreticulin; D002352:Carrier Proteins / Q000032:analysis; D002417:Cattle; D002460:Cell Line; D002846:Chromatography, Affinity; D003208:Concanavalin A / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D000091342:Endoplasmic Reticulum Chaperone BiP; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D006360:Heat-Shock Proteins; D007668:Kidney / Q000166:cytology* / Q000378:metabolism*; D008715:Methionine / Q000032:analysis / Q000378:metabolism; D018832:Molecular Chaperones; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012261:Ribonucleoproteins / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013462:Sulfur Radioisotopes
D000596:Amino Acids; D002135:Calcium-Binding Proteins; D037282:Calreticulin; D002352:Carrier Proteins; D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D012261:Ribonucleoproteins; D013462:Sulfur Radioisotopes; D003208:Concanavalin A; D008715:Methionine