The C-terminal region of alpha' subunit of soybean beta-conglycinin contains two types of vacuolar sorting determinants.
In maturing seed cells, proteins that accumulate in the protein storage vacuoles (PSVs) are synthesized on the endoplasmic reticulum (ER) and transported by vesicles to the PSVs. Vacuolar sorting determinants (VSDs) which are usually amino acid sequences of short or moderate length direct the proteins to this pathway. VSDs identified so far are classified into two types: sequence specific VSDs (ssVSDs) and C-terminal VSDs (ctVSDs). We previously demonstrated that VSDs of alpha' and beta subunits of beta-conglycinin, one of major storage proteins of soybean (Glycine max), reside in the C-terminal ten amino acids. Here we show that both types of VSDs coexist within this region of the alpha' subunit. Although ctVSDs can function only at the very C-termini of proteins, the C-terminal ten amino acids of alpha' subunit directed green fluorescent protein (GFP) to the PSVs even when they were placed at the N-terminus of GFP, indicating that an ssVSD resides in the sequence. By mutation analysis, it was found that the core sequence of the ssVSD is Ser-Ile-Leu (fifth to seventh residues counted from the C-terminus) which is conserved in the alpha and beta subunits and some vicilin-like proteins. On the other hand, the sequence composed of the C-terminal three amino acids (AFY) directed GFP to the PSVs when it was placed at the C-terminus of GFP, though the function as a VSD was disrupted at the N-terminus of GFP, indicating that the AFY sequence is a ctVSD.
Nishizawa
Keito
K
Laboratory of Food Quality Design and Development, Graduate School of Agriculture, Kyoto University, Uji, Kyoto 611-0011, Japan.
Maruyama
Nobuyuki
N
Utsumi
Shigeru
S
D000595:Amino Acid Sequence; D052179:Antigens, Plant; D001483:Base Sequence; D017931:DNA Primers; D018506:Gene Expression Regulation, Plant; D005916:Globulins / Q000737:chemistry* / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D055314:Seed Storage Proteins; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D030262:Soybean Proteins / Q000737:chemistry* / Q000378:metabolism*; D013025:Glycine max / Q000235:genetics / Q000502:physiology*; D014617:Vacuoles / Q000502:physiology
D052179:Antigens, Plant; D017931:DNA Primers; D005916:Globulins; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; D055314:Seed Storage Proteins; D030262:Soybean Proteins; C013894:beta-conglycinin protein, Glycine max; D049452:Green Fluorescent Proteins