Kinetic studies of the inhibitory effects of propeptides subtilisin BPN' and Carlsberg to bacterial serine proteases.
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.
Huang
H W
HW
Institute of Biological Chemistry, Academia Sinica, National Taiwan University, Republic of China.
Chen
W C
WC
Wu
C Y
CY
Yu
H C
HC
Lin
W Y
WY
Chen
S T
ST
Wang
K T
KT
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
D000596:Amino Acids; D001426:Bacterial Proteins; D009842:Oligopeptides; D011498:Protein Precursors; D015842:Serine Proteinase Inhibitors; D013381:Subtilisins