Human mast cell tryptase attenuates the vasodilator activity of calcitonin gene-related peptide.
Calcitonin gene-related peptide (CGRP) is localized in and released from sensory nerves. It is a potent and long acting vasodilator which has been suggested to play a role in the control of blood flow. Using HPLC and trichloroacetic acid precipitation techniques, we have examined the ability of human mast cell lysates and a purified preparation of mast cell tryptase to degrade CGRP. We found that CGRP is effectively cleaved by tryptase (Km = 6.8 x 10(-6) mol/L at 37 degrees). Enzymatic activity was inhibited by antipain, leupeptin, N-alpha-p-tosyl-L-lysine chloromethyl ketone, benzamidine or aprotinin, but not by soybean trypsin inhibitor or N-tosyl-L-phenylalanine chloromethyl ketone. The degradation of CGRP by lysates of purified skin mast cells showed a similar pattern of inhibition suggesting that tryptase may be the major enzyme involved. The activity of tryptase was not affected by the presence of heparin. Incubation of CGRP with tryptase resulted in a loss of its vasodilator activity as observed by intravital microscopy of the hamster cheek pouch microvasculature. CGRP preincubated with tryptase failed to relax arterioles when added topically. It is suggested that the catalysis of CGRP by tryptase could represent an important means by which the activity of this neuropeptide is regulated in vivo.
Walls
A F
AF
Immunopharmacology Group, Southampton General Hospital, Southampton, U.K.
Brain
S D
SD
Desai
A
A
Jose
P J
PJ
Hawkings
E
E
Church
M K
MK
Williams
T J
TJ
D015740:Calcitonin Gene-Related Peptide / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008407:Mast Cells / Q000187:drug effects / Q000201:enzymology*; D010447:Peptide Hydrolases / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology; D012867:Skin / Q000201:enzymology; D014665:Vasodilator Agents / Q000378:metabolism
D011480:Protease Inhibitors; D014665:Vasodilator Agents; D010447:Peptide Hydrolases; C023073:tosylarginine methyl ester hydrolase; D015740:Calcitonin Gene-Related Peptide