Differences in the P1' substrate specificities of pepsin A and chymosin.
Porcine pepsin A and bovine chymosin are typical models of aspartic proteinases. The hydrolytic specificities of these proteinases, along with those of human pepsin A and monkey chymosin, were investigated with 29 peptide substrates that included various P1' variants of seven parent peptides. From these peptides, AFPLEF downward arrow FREL was preferred by pepsin A and chymosin, while its P1' variant, AFPLEF downward arrow EREL was preferred by bovine chymosin. Porcine and human pepsin A showed similar hydrolytic specificities, strongly preferring a hydrophobic/aromatic residue at P1' of any type of peptide. This specificity is well explained by the very hydrophobic nature of the S1' subsite that consists of Tyr(189), Ile(213), Ile(300), Met(289), Val/Leu(291) and Leu(298). The first three residues are well conserved in pepsin family enzymes. Although bovine and monkey chymosin showed similar P1' specificity, bovine chymosin preferred peptides having Glu at P1', while monkey chymosin preferred peptides having Lys at P1'. The dual characteristics of chymosin are due to the occurrence of polar/charged residues in the S1' subsite, such as Glu/Asp(289), Gln(298) and Lys/Gln(299), which are different from the S1' subsite of pepsin A. Molecular models suggest that Glu in position 289 of bovine chymosin and Asp in position 289 of monkey chymosin are responsible for the difference in P1' specificities between the chymosins.
Kageyama
Hakuto
H
School of Science, Biological Science, Nagoya University, Japan. hakuto_kageyama@ajinomoto.com
Ueda
Hiroshi
H
Tezuka
Takafumi
T
Ogasawara
Asami
A
Narita
Yuichi
Y
Kageyama
Takashi
T
Ichinose
Masao
M
D000818:Animals; D012085:Chymosin / Q000378:metabolism*; D000882:Haplorhini; D006801:Humans; D007700:Kinetics; D056004:Molecular Dynamics Simulation; D010434:Pepsin A / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity
D010455:Peptides; D010434:Pepsin A; D012085:Chymosin