The thiol proteinases from the latex of Carica papaya L. IV. Proteolytic specificities of chymopapain and papaya proteinase omega determined by digestion of alpha-globin chains.
The proteolytic specificities of chymopapain and papaya proteinase omega were investigated by using the alpha-chains of manatee and mole haemoglobin, whose primary structures are known, as substrates. The resulting peptides from each enzymatic cleavage were isolated by gel filtration on Sephadex G-25, followed by reversed-phase HPLC of the separated peaks and, in some cases, further purified by preparative thin-layer electrophoresis. The purified peptides were then identified on the basis of their amino-acid composition. The proteolytic specificities of chymopapain and papaya proteinase omega, deduced from the experimental cleavage patterns, are compared to that of papain. As in the case of papain, the specificity-determining factor is the amino-acid residue of the substrate that will be bound in subsite S2 (the next but one from the scissible bond). Aromatic residues in this position, preferred by papain, are not important for chymopapain and papaya proteinase omega. Cleavages preferentially occur when S2 is occupied by leucine, valine or threonine. For chymopapain, proline in position S2 also causes cleavage.
Jacquet
A
A
Laboratoire de Chimie Générale I, Université de Bruxelles.
Kleinschmidt
T
T
Dubois
T
T
Schnek
A G
AG
Looze
Y
Y
Braunitzer
G
G
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002917:Chymopapain / Q000378:metabolism*; D004586:Electrophoresis; D010450:Endopeptidases / Q000378:metabolism*; D005914:Globins / Q000378:metabolism*; D006454:Hemoglobins / Q000378:metabolism; D007840:Latex / Q000032:analysis; D008969:Molecular Sequence Data; D008971:Moles; D010944:Plants / Q000201:enzymology*; D013379:Substrate Specificity
D000596:Amino Acids; D006454:Hemoglobins; D007840:Latex; D005914:Globins; D010450:Endopeptidases; D002917:Chymopapain