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The 2.2 A Crystal Structure Of Human Chymase In Complex With Succinyl-Ala-Ala-Pro-Phe-chloromethylketone: Structural Explanation For Its Dipeptidyl Carboxypeptidase Specificity.

P. J. Pereira, Z. Wang, H. Rubin, R. Huber, W. Bode, N. Schechter, S. Strobl
Published 1999 · Chemistry, Medicine

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Human chymase (HC) is a chymotrypsin-like serine proteinase expressed by mast cells. The 2.2 A crystal structure of HC complexed to the peptidyl inhibitor, succinyl-Ala-Ala-Pro-Phe-chloromethylketone (CMK), was solved and refined to a crystallographic R-factor of 18.4 %. The HC structure exhibits the typical folding pattern of a chymotrypsin-like serine proteinase, and shows particularly similarity to rat chymase 2 (rat mast cell proteinase II) and human cathepsin G. The peptidyl-CMK inhibitor is covalently bound to the active-site residues Ser195 and His57; the peptidyl moiety juxtaposes the S1 entrance frame segment 214-217 by forming a short antiparallel beta-sheet. HC is a highly efficient angiotensin-converting enzyme. Modeling of the chymase-angiotensin I interaction guided by the geometry of the bound chloromethylketone inhibitor indicates that the extended substrate binding site contains features that may generate the dipeptidyl carboxypeptidase-like activity needed for efficient cleavage and activation of the hormone. The C-terminal carboxylate group of angiotensin I docked into the active-site cleft, with the last two residues extending beyond the active site, is perfectly localized to make a favorable hydrogen bond and salt bridge with the amide nitrogen of the Lys40-Phe41 peptide bond and with the epsilon-ammonium group of the Lys40 side-chain. This amide positioning is unique to the chymase-related proteinases, and only chymases from primates possess a Lys residue at position 40. Thus, the structure conveniently explains the preferred conversion of angiotensin I to angiotensin II by human chymase.
This paper references
The CCP4 suite: programs for protein crystallography.
Collaborative Computational (1994)
Human Prochymase Activation
M. Murakami (1995)
Natural protein proteinase inhibitors and their interaction with proteinases.
W. Bode (1992)
Location of disulphide bridges by diagonal paper electrophoresis. The disulphide bridges of bovine chymotrypsinogen A.
J. R. Brown (1966)
Inhibitors of human heart chymase based on a peptide library.
M. Bastos (1995)
Mast Cell Proteases in Immunology and Biology
G. Caughey (1995)
The refined 2.15 A X‐ray crystal structure of human liver cathepsin B: the structural basis for its specificity.
D. Musil (1991)
Substance P regulates the vasodilator activity of calcitonin gene-related peptide
S. Brain (1988)
Mast cell tryptase and chymase reverse airway smooth muscle relaxation induced by vasoactive intestinal peptide in the ferret.
G. Franconi (1989)
Crystallographic phasing and refinement of macromolecules: Current Opinion in Structural Biology 1991, 1:1016–1022
A. Brunger (1991)
Human β-tryptase is a ring-like tetramer with active sites facing a central pore
P. Pereira (1998)
Rapid and specific conversion of precursor interleukin 1 beta (IL-1 beta) to an active IL-1 species by human mast cell chymase
H. Mizutani (1991)
Production of active recombinant human chymase from a construct containing the enterokinase cleavage site of trypsinogen in place of the native propeptide sequence.
Z. Wang (1995)
ALSCRIPT: a tool to format multiple sequence alignments.
G. Barton (1993)
Rapid conversion of angiotensin I to angiotensin II by neutrophil and mast cell proteinases.
C. Reilly (1982)
Two types of human mast cells that have distinct neutral protease compositions.
A. A. Irani (1986)
The 1.8 A crystal structure of human cathepsin G in complex with Suc‐Val‐Pro‐PheP‐(OPh)2: a Janus‐faced proteinase with two opposite specificities.
P. Hof (1996)
Crystal structure of the bifunctional soybean Bowman-Birk inhibitor at 0.28-nm resolution. Structural peculiarities in a folded protein conformation.
R. H. Voss (1996)
The refined crystal structure of bovine beta-trypsin at 1.8 A resolution. II. Crystallographic refinement, calcium binding site, benzamidine binding site and active site at pH 7.0.
W. Bode (1975)
Mast cell chymase. A potent secretagogue for airway gland serous cells.
C. Sommerhoff (1989)
SETOR: hardware-lighted three-dimensional solid model representations of macromolecules.
S. V. Evans (1993)
The structure of rat mast cell protease II at 1.9-A resolution.
S. Remington (1984)
Substrate specificity of the chymotrypsin-like protease in secretory granules isolated from rat mast cells.
H. Le Trong (1987)
Genomic organization and chromosomal localization of the human cathepsin G gene.
P. A. Hohn (1989)
Crystal structure of phenylmethanesulfonyl fluoride-treated human chymase at 1.9 A.
M. Mcgrath (1997)
Bovine chymotrypsinogen A X-ray crystal structure analysis and refinement of a new crystal form at 1.8 A resolution.
D. Wang (1985)
Angiotensin II-Forming Activity in a Reconstructed Ancestral Chymase
U. Chandrasekharan (1996)
AMoRe: an automated package for molecular replacement
J. Navaza (1994)
Mast cells at sites of cartilage erosion in the rheumatoid joint.
M. Bromley (1984)
Angiotensin I conversion by human and rat chymotryptic proteinases.
B. Wintroub (1984)
Recombinant Expression of Human Mast Cell Proteases Chymase and Tryptase
Z. Wang (1998)
Mammalian chymotrypsin-like enzymes. Comparative reactivities of rat mast cell proteases, human and dog skin chymases, and human cathepsin G with peptide 4-nitroanilide substrates and with peptide chloromethyl ketone and sulfonyl fluoride inhibitors.
J. Powers (1985)
Identification of a highly specific chymase as the major angiotensin II-forming enzyme in the human heart.
H. Urata (1990)
Soybean Bowman-Birk protease inhibitor is a highly effective inhibitor of human mast cell chymase.
J. Ware (1997)
Accurate Bond and Angle Parameters for X-ray Protein Structure Refinement
R. Engh (1991)
Substance P and vasoactive intestinal peptide degradation by mast cell tryptase and chymase.
G. Caughey (1988)
Multiple determinants for the high substrate specificity of an angiotensin II-forming chymase from the human heart.
A. Kinoshita (1991)
PROCHECK: a program to check the stereochemical quality of protein structures
R. Laskowski (1993)
On the size of the active site in proteases. I. Papain.
I. Schechter (1967)
Distinct Multisite Synergistic Interactions Determine Substrate Specificities of Human Chymase and Rat Chymase-1 for Angiotensin II Formation and Degradation*
S. Sanker (1997)
Activation of human interstitial procollagenase through direct cleavage of the Leu83-Thr84 bond by mast cell chymase.
J. Saarinen (1994)
The refinement and the structure of the dimer of alpha-chymotrypsin at 1.67-A resolution.
R. Blevins (1985)

This paper is referenced by
Extended substrate specificity of opossum chymase--implications for the origin of mast cell chymases.
J. Reimer (2008)
Non-peptidic inhibitors of human chymase. Synthesis, structure-activity relationships, and pharmacokinetic profiles of a series of 5-amino-6-oxo-1,6-dihydropyrimidine-containing trifluoromethyl ketones.
F. Akahoshi (2001)
Quantum chemical study on the deacylation step of human chymase
Y. Hirano (2005)
Inhibitors of human mast cell serine proteases and potential therapeutic applications
Kenneth D. Rice (1999)
Evidence for diversity of substrate specificity among members of the chymase family of serine proteases
S. Solivan (2002)
Cloning and Molecular Modeling of Duodenase with Respect to Evolution of Substrate Specificity within Mammalian Serine Proteases That Have Lost a Conserved Active-Site Disulfide Bond
T. S. Zamolodchikova (2005)
Generation and characterization of new monoclonal antibodies against human chymase.
Y. Kunori (2004)
The human mast cell tryptase tetramer: a fascinating riddle solved by structure.
C. Sommerhoff (2000)
Cleavage Specificity of Mast Cell Chymases
M. Andersson (2008)
Lys40 but not Arg143 influences selectivity of angiotensin conversion by human α-chymase
D. Muilenburg (2002)
Therapeutic targeting of cathepsin C: from pathophysiology to treatment
B. Korkmaz (2018)
Inhibition of human chymase by 2-amino-3,1-benzoxazin-4-ones.
U. Neumann (2001)
The extended substrate specificity of the human mast cell chymase reveals a serine protease with well-defined substrate recognition profile.
M. Andersson (2009)
Arg143 and Lys192 of the human mast cell chymase mediate the preference for acidic amino acids in position P2′ of substrates
M. Andersson (2010)
Molecular Cloning and Sequencing of the cDNA for Rat Mesenteric Arterial Bed Elastase-2, an Angiotensin II–Forming Enzyme
C. Santos (2002)
The three-dimensional structure of human granzyme B compared to caspase-3, key mediators of cell death with cleavage specificity for aspartic acid in P1.
J. Rotonda (2001)
Inhibitory mechanism of daphnodorins for human chymase.
M. Sakaguchi (2001)
Expressão e caracterização das quimases recombinantes específicas de mastócitos de camundongos (mMCP4 e 5)
Programa de Pós-graduação (2014)
Expression of recombinant human mast cell chymase with Asn-linked glycans in glycoengineered Pichia pastoris.
Eliot T Smith (2014)
Expression, Purification, and Characterization of the Mast Cell Proteases Chymase and Cathepsin G.
B. E. Lockhart (2008)
Discriminating between the activities of human cathepsin G and chymase using fluorogenic substrates
B. Korkmaz (2011)
Chymase inhibitor-sensitive synthesis of endothelin-1 (1-31) by recombinant mouse mast cell protease 4 and human chymase.
W. Semaan (2015)
Chapter 18 – Structure and Function of Human Chymase
N. Schechter (2000)
Structure-activity relationship of benzo[b]thiophene-2-sulfonamide derivatives as novel human chymase inhibitors.
Hidekazu Masaki (2003)
An assessment of protein-ligand binding site polarizability.
A. Nayeem (2003)
Design, synthesis and pharmacological evaluation of 3-benzylazetidine-2-one-based human chymase inhibitors.
Y. Aoyama (2001)
Rodent α‐chymases are elastase‐like proteases
Y. Kunori (2002)
Mouse mast cell protease-1 cleaves angiotensin I to form angiotensin II.
Kayo Saito (2003)
Highly efficient inhibition of human chymase by alpha(2)-macroglobulin.
M. Walter (1999)
Mutations in Arg143 and Lys192 of the Human Mast Cell Chymase Markedly Affect the Activity of Five Potent Human Chymase Inhibitors
P. Ahooghalandari (2013)
A Pulmonary Perspective on GASPIDs: Granule-Associated Serine Peptidases of Immune Defense.
G. Caughey (2006)
Mast cell proteases.
G. Pejler (2007)
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