title: Kinetic studies of the inhibitory effects of propeptides subtilisin BPN' and Carlsberg to bacterial serine proteases.
abstract: The propeptides of bacterial subtilisin BPN' and Carlsberg were synthesized to investigate their inhibitory function on the enzymes. Kinetically, pro-BPN' inhibits the proteolytic activities of subtilisin BPN' and Carlsberg separately in a slow binding mode. Pro-Carlsberg behaves as a typical rapid equilibrium competitive inhibitor for these two proteases. Functionally, pro-Carlsberg inhibits the subtilisins with moderate selectivity. The inhibition constant Ki of pro-BPN' to subtilisin BPN' is 5.0 nM, and 6.1 nM to subtilisin Carlsberg. The on-rate of pro-BPN' to subtilisin BPN' is 5.8 x 10(5) M(-1)s(-1), and the off-rate 2.9 x 10(-3) s(-1). Similarly, the on-rate of pro-BPN' to subtilisin Carlsberg is 2.2 x 10(5) M(-1)s(-1), and the off-rate 1.3 x 10(-3) s(-1). On the other hand, the Ki of pro-Carlsberg to subtilisin BPN' gives 1.3 x 10(2) nM, and 88 nM to subtilisin Carlsberg. Based on the key features of the interactions between pro-BPN' and subtilisin from X-ray crystallographic results (Gallagher et al., 1995), the correlation between the sequence of subtilisin propeptides and their inhibition abilities on the proteases are compared and discussed.
authors.affiliation: Institute of Biological Chemistry, Academia Sinica, National Taiwan University, Republic of China.
authors.forename: H W
authors.initials: HW
authors.lastname: Huang
authors.forename: W C
authors.initials: WC
authors.lastname: Chen
authors.forename: C Y
authors.initials: CY
authors.lastname: Wu
authors.forename: H C
authors.initials: HC
authors.lastname: Yu
authors.forename: W Y
authors.initials: WY
authors.lastname: Lin
authors.forename: S T
authors.initials: ST
authors.lastname: Chen
authors.forename: K T
authors.initials: KT
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000378:metabolism; D009994:Osmolar Concentration; D011498:Protein Precursors / Q000737:chemistry* / Q000378:metabolism; D015842:Serine Proteinase Inhibitors / Q000737:chemistry* / Q000378:metabolism; D013381:Subtilisins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D000596:Amino Acids; D001426:Bacterial Proteins; D009842:Oligopeptides; D011498:Protein Precursors; D015842:Serine Proteinase Inhibitors; D013381:Subtilisins