Limited proteolysis regulates massive degradation of glycinin, storage 11S globulin from soybean seeds: an in vitro model.
The time course of glycinin hydrolysis by papain was followed using densitometry of SDS-PAGE patterns, quantification of the residual protein and determination of its molecular mass by gel filtration, and by other appropriate methods. The hydrolysis occurs in two steps. In the first step, a limited proteolysis was observed consisting of a gradual detachment of the α-chain C-terminal sequence region, leading to the formation of glycinin-P, a relatively stable proteolysis product retaining the primordial hexameric structure. Glycinin-P was found to be composed of the intact β-chains covalently bound with the C-terminally truncated α-chains lacking the helix domain, strand J', and the C-terminal disordered region. Glycinin-P is further hydrolyzed in the second step exclusively by a one-by-one mechanism. Comparison of the kinetics of the limited and one-by-one proteolyses analyzed separately indicated that the decrease of protein concentration by 24-25% in the first step occurs almost exclusively due to the decrease of the molecular mass of the residual protein. Thus, the onset of the one-by-one proteolysis is delayed, suggesting a regulatory role of the preceding limited proteolysis in the subsequent massive degradation of glycinin. Probable structural alterations of glycinin generated by this limited proteolysis are discussed.
Laboratory of Plant Biochemistry, Moldova State University, Mateevici str. 60, MD-2009 Chişinău, Republic of Moldova.
Andrei
A
Shutov
Angela
A
Rudakova
Sergei
S
Rudakov
Irina
I
Kakhovskaya
Anna
A
Schallau
Nobuyuki
N
Maruyama
Karl
K
Wilson
D005916:Globulins / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D011487:Protein Conformation; D059748:Proteolysis; D055314:Seed Storage Proteins / Q000378:metabolism*; D012639:Seeds / Q000378:metabolism*; D030262:Soybean Proteins / Q000737:chemistry / Q000378:metabolism*; D013025:Glycine max / Q000378:metabolism*
D005916:Globulins; D055314:Seed Storage Proteins; D030262:Soybean Proteins; C011474:glycinin