Calculation of subunit stoichiometry of large, multisubunit proteins from amino acid compositions.
The subunit stoichiometry of a large, multisubunit protein can be determined from the molar amino acid compositions (i amino acids) of the protein and its subunits. The number of copies of the subunits (1, 2, ... j) is calculated by solving all possible combinations of simultaneous equations in j unknowns (i!/j!(i - j)!). Calculations carried out using the published amino acid compositions determined by analysis and the compositions calculated from the sequences for two proteins of known stoichiometry provided the following results: Escherichia coli aspartate transcarbamoylase (R6C6, Mr = 307.5 kDa), R = 5.6 to 6.6 and C = 5.8 to 6.3, and spinach ribulose-bisphosphate carboxylase (L8S8, Mr = 535 kDa), L = 7.3 to 9.1 and S = 5.6 to 10.6. Calculations were also carried out with the amino acid compositions of two much larger proteins, the E. coli pyruvate dehydrogenase complex, Mr = 5280 kDa, subunits E1 (99.5 kDa), E2 (66 kDa), and E3 (50.6 kDa), and the extracellular hemoglobin of Lumbricus terrestris, Mr = 3760 kDa, subunits M (17 kDa), D1 (31 kDa), D2 (37 kDa), and T (51 kDa); the results for PDHase were E1 = 20 to 24, E2 = 18 to 31, E3 = 21 to 33 and those for Lumbricus hemoglobin were M = 34 to 46, D1 = 13 to 19, D2 = 13 to 18, and T = 34 to 36. Although the sample standard deviations of the mean values are generally high, the proposed method works surprisingly well for the two smaller proteins and provides physically reasonable results for the two larger proteins.
Kapp
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Enrico Fermi Institute, University of Chicago, Illinois 60637.
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Vinogradov
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D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001221:Aspartate Carbamoyltransferase / Q000032:analysis; D006454:Hemoglobins / Q000032:analysis*; D008433:Mathematics; D015394:Molecular Structure; D011506:Proteins / Q000032:analysis*; D012273:Ribulose-Bisphosphate Carboxylase / Q000032:analysis
D000596:Amino Acids; D006454:Hemoglobins; D011506:Proteins; D001221:Aspartate Carbamoyltransferase; D012273:Ribulose-Bisphosphate Carboxylase