A study was conducted to investigate the amino acid (AA) composition and the susceptibility to in vitro proteolysis (pepsin, 120 min and pancreatin, 240 min) of a collection of purified phaseolins (n = 43) in unheated or heat-treated form. The AA composition of phaseolin varied little across bean varieties. At 360 min of in vitro proteolysis, the degree of hydrolysis varied from 11 to 27% for unheated and from 57 to 96% for heated phaseolins (P < 0.001). Heat treatment markedly increased the susceptibility of phaseolin to proteolysis (P < 0.001). The AA scores (AAS) and the protein digestibility corrected for AAS indicated S-containing AA as the limiting AA (39 ± 3 and 30 ± 5%, respectively). In conclusion, susceptibility to proteolysis of heat-treated phaseolin rather than its AA composition affects the nutritional value of phaseolin estimated in vitro. Therefore, it should be the criterion of choice in breeding programs aimed at improving the nutritional value of common beans for humans.