A rapid increase in the level of binding protein (BiP) is accompanied by synthesis and degradation of storage proteins in pumpkin cotyledons.
The binding protein (BiP) has been implicated in cotranslational folding of nascent polypeptides, and in the recognition and disposal of aberrant polypeptides. To elucidate the involvement of BiP in the biosynthesis of vacuolar proteins, we have characterized the protein in pumpkin cotyledons during seed maturation and seedling growth. Isolated microsomes from maturing pumpkin cotyledons contained a significant amount of BiP, protein-disulfide isomerase and calreticulin. We have purified a 70-kDa protein; sequences of the N-terminus and internal fragments of this protein exhibited a high identity to the sequence of soybean. Immunoblot analysis with specific antibodies raised against the purified BiP showed that the amount of BiP in a cotyledon increased markedly at the middle stages and then decreased. The increase was accompanied by the synthesis of storage proteins and the development of the endoplasmic reticulum in the cotyledons at the middle stage of seed maturation. Most of these storage proteins degraded dramatically between 2 and 5 days after seed germination, and the degradation was also accompanied by a rapid increase in the level of BiP. Subcellular fractionation of the 4-day-old cotyledons showed a high accumulation of BiP in the endoplasmic reticulum. It is possible that BiP might be involved in the synthesis of seed storage proteins during maturation and in the synthesis of hydrolytic enzymes responsible for the degradation of the storage proteins during seed germination.
Hatano
K
K
Department of Cell Biology, National Institute for Basic Biology, Okazaki, Japan.
Shimada
T
T
Hiraiwa
N
N
Nishimura
M
M
Hara-Nishimura
I
I
D000595:Amino Acid Sequence; D002135:Calcium-Binding Proteins / Q000378:metabolism; D037282:Calreticulin; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D018548:Cotyledon; D003624:Darkness; D006360:Heat-Shock Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D007535:Isomerases / Q000378:metabolism; D008027:Light; D008854:Microscopy, Electron; D008861:Microsomes / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry; D010940:Plant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D019704:Protein Disulfide-Isomerases; D012261:Ribonucleoproteins / Q000378:metabolism; D014675:Vegetables / Q000254:growth & development / Q000378:metabolism* / Q000648:ultrastructure
D002135:Calcium-Binding Proteins; D037282:Calreticulin; D002352:Carrier Proteins; D006360:Heat-Shock Proteins; D010446:Peptide Fragments; D010940:Plant Proteins; D012261:Ribonucleoproteins; D007535:Isomerases; D019704:Protein Disulfide-Isomerases