A 28-kilodalton pod storage protein of French bean plants. Purification, characterization, and primary structure.
When French bean (Phaseolus vulgaris) plants were depodded in the early stages of fruit development, relative levels of a specific protein with a relative molecular weight of 28,000 were enhanced in the young pods that formed later. The protein, designated pod storage protein (PSP), was purified from extracts of newly formed pods from plants that had been previously depodded four times at intervals of 2 weeks. Two-dimensional polyacrylamide gel electrophoresis showed the presence of three forms (designated A, B, and C) of PSP with identical electrophoretic mobilities but different charges. The molecular mass of native PSP was estimated by gel filtration to be 67 kD; therefore, the protein was most likely present as a dimer. The antisera raised against forms A and C were crossreactive with each other. Form B lacked the N-terminal alanine of forms A and C. An expression library from French bean pods was screened using the antiserum against form A, and a full-length cDNA clone was isolated. The cDNA insert included 765 bp potentially encoding a polypeptide with 255 amino acid residues (and a calculated molecular mass of 28,854 D). The amino acid sequence deduced from the PSP cDNA had 65 to 71% identity with soybean (Glycine max) vegetative storage protein sequences (P.E. Staswick [1988] Plant Physiol 87: 250-254; and Correction [1989] Plant Physiol 89: 717). Genomic Southern blot analysis suggested that PSP is derived from a single-copy gene.
Zhong
P Y
PY
Department of Biology, Tokyo Metropolitan University, Japan.
Tanaka
T
T
Yamauchi
D
D
Minamikawa
T
T
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D007887:Fabaceae / Q000737:chemistry / Q000235:genetics*; D018628:Gene Dosage; D017343:Genes, Plant; D008969:Molecular Sequence Data; D018515:Plant Leaves / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics*; D018520:Plant Shoots / Q000737:chemistry / Q000235:genetics*; D010946:Plants, Medicinal; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D018749:RNA, Plant / Q000032:analysis / Q000235:genetics; D055314:Seed Storage Proteins; D017422:Sequence Analysis, DNA; D014018:Tissue Distribution
D018076:DNA, Complementary; C104899:PSP protein, Phaseolus vulgaris; D010940:Plant Proteins; D012333:RNA, Messenger; D018749:RNA, Plant; D055314:Seed Storage Proteins