The three-dimensional structure of recombinant leech-derived tryptase inhibitor in complex with trypsin. Implications for the structure of human mast cell tryptase and its inhibition.
The x-ray crystal structure of recombinant leech-derived tryptase inhibitor (rLDTI) has been solved to a resolution of 1.9 A in complex with porcine trypsin. The nonclassical Kazal-type inhibitor exhibits the same overall architecture as that observed in solution and in rhodniin. The complex reveals structural aspects of the mast cell proteinase tryptase. The conformation of the binding region of rLDTI suggests that tryptase has a restricted active site cleft. The basic amino terminus of rLDTI, apparently flexible from previous NMR measurements, approaches the 148-loop of trypsin. This loop has an acidic equivalent in tryptase, suggesting that the basic amino terminus could make favorable electrostatic interactions with the tryptase molecule. A series of rLDTI variants constructed to probe this hypothesis confirmed that the amino-terminal Lys-Lys sequence plays a role in inhibition of human lung tryptase but not of trypsin or chymotrypsin. The location of such an acidic surface patch is in accordance with the known low molecular weight inhibitors of tryptase.
Stubbs
M T
MT
Max-Planck-Institut für Biochemie, Am Klopferspitz, D-82152 Martinsried bei München, Germany.
Morenweiser
R
R
Stürzebecher
J
J
Bauer
M
M
Bode
W
W
Huber
R
R
Piechottka
G P
GP
Matschiner
G
G
Sommerhoff
C P
CP
Fritz
H
H
Auerswald
E A
EA
D000595:Amino Acid Sequence; D000818:Animals; D053818:Chymases; D018360:Crystallography, X-Ray; D006801:Humans; D007865:Leeches; D008168:Lung / Q000201:enzymology; D008407:Mast Cells / Q000201:enzymology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000737:chemistry / Q000378:metabolism*; D013552:Swine; D014357:Trypsin / Q000378:metabolism*; D053802:Tryptases
D011506:Proteins; D015842:Serine Proteinase Inhibitors; C090901:leech-derived tryptase inhibitor C; D012697:Serine Endopeptidases; C083294:chymase 2; D053818:Chymases; D014357:Trypsin; D053802:Tryptases