Adenosine 5'-triphosphate is required for the assembly of 11S seed proglobulins in vitro.
Seed protein proglobulins were synthesized from cDNAs in reticulocyte lysates. Most proglobulins were recovered as trimers when translation rates were low, but mostly monomers were recovered at high translation rates. The prevalence of monomers was accompanied by elevated amounts of insoluble protein recovered at the bottom of sucrose density gradients. Apyrase treatment of translation mixtures after synthesis, but before significant assembly occurred, drastically reduced trimer assembly and increased the proportion of insoluble aggregate. These observations indicated that ATP is required for protein folding and/or trimer assembly. The appearance of insoluble aggregated protein when rates of synthesis were elevated or when ATP was absent suggested that protein misfolding had occurred. Trimer assembly was stimulated when wheat germ translation mixtures defective in supporting efficient trimer assembly were supplemented with fractions isolated from endoplasmic reticula of developing pea (Pisum sativum) seeds. Molecular chaperones are likely involved in folding and/or assembly of proglobulin trimers both in reticulocyte lysates and in seeds. Consistent with this hypothesis, trimer formation was reduced when carboxymethylated bovine albumin and alpha-casein, considered to mimic proteins with extended chain and molten globular conformations and thereby compete for Hsp70- and Hsp60-type molecular chaperones, respectively, were introduced into translation mixtures.
Nam
Y W
YW
United States Department of Agriculture, Agricultural Research Service, West Lafayette, Indiana, USA.
Jung
R
R
Nielsen
N C
NC
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D001081:Apyrase / Q000378:metabolism; D002364:Caseins / Q000494:pharmacology; D002417:Cattle; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005916:Globulins / Q000096:biosynthesis* / Q000737:chemistry; D046911:Macromolecular Substances; D018832:Molecular Chaperones / Q000378:metabolism; D018532:Pisum sativum / Q000378:metabolism*; D010940:Plant Proteins / Q000096:biosynthesis* / Q000737:chemistry; D014176:Protein Biosynthesis; D017510:Protein Folding; D011498:Protein Precursors / Q000096:biosynthesis* / Q000737:chemistry; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism; D012639:Seeds / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D014158:Transcription, Genetic; D014908:Triticum / Q000378:metabolism
D002364:Caseins; D005916:Globulins; D046911:Macromolecular Substances; D018832:Molecular Chaperones; D010940:Plant Proteins; D011498:Protein Precursors; D012710:Serum Albumin, Bovine; D000255:Adenosine Triphosphate; D001081:Apyrase