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α2-Macroglobulin Capture Allows Detection Of Mast Cell Chymase In Serum And Creates A Reservoir Of Angiotensin II-Generating Activity1
W. Raymond, S. Su, A. Makarova, T. M. Wilson, M. Carter, D. Metcalfe, G. Caughey
Published 2009 · Biology, Medicine
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Human chymase is a highly efficient angiotensin II-generating serine peptidase expressed by mast cells. When secreted from degranulating cells, it can interact with a variety of circulating antipeptidases, but is mostly captured by α2-macroglobulin, which sequesters peptidases in a cage-like structure that precludes interactions with large protein substrates and inhibitors, like serpins. The present work shows that α2-macroglobulin-bound chymase remains accessible to small substrates, including angiotensin I, with activity in serum that is stable with prolonged incubation. We used α2-macroglobulin capture to develop a sensitive, microtiter plate-based assay for serum chymase, assisted by a novel substrate synthesized based on results of combinatorial screening of peptide substrates. The substrate has low background hydrolysis in serum and is chymase-selective, with minimal cleavage by the chymotryptic peptidases cathepsin G and chymotrypsin. The assay detects activity in chymase-spiked serum with a threshold of ∼1 pM (30 pg/ml), and reveals native chymase activity in serum of most subjects with systemic mastocytosis. α2-Macroglobulin-bound chymase generates angiotensin II in chymase-spiked serum, and it appears in native serum as chymostatin-inhibited activity, which can exceed activity of captopril-sensitive angiotensin-converting enzyme. These findings suggest that chymase bound to α2-macroglobulin is active, that the complex is an angiotensin-converting enzyme inhibitor-resistant reservoir of angiotensin II-generating activity, and that α2-macroglobulin capture may be exploited in assessing systemic release of secreted peptidases.
This paper references
α2“Macroglobulin, a multifunctional binding protein with targeting characteristics
W. Borth (1992)
Lys40 but not Arg143 influences selectivity of angiotensin conversion by human α-chymase
D. Muilenburg (2002)
Tryptase levels as an indicator of mast-cell activation in systemic anaphylaxis and mastocytosis.
L. Schwartz (1987)
Chymase expressing bone marrow mast cells in mastocytosis and myelodysplastic syndromes: an immunohistochemical and morphometric study
H.-P. Horny (2003)
Highly efficient inhibition of human chymase by 2-macroglobulin
M. Walter (1999)
Effect of sex and haplotype on plasma tryptase levels in healthy adults.
H. Min (2004)
Pathological roles of angiotensin II produced by mast cell chymase and the effects of chymase inhibition in animal models.
M. Miyazaki (2006)
Tranilast suppresses vascular chymase expression and neointima formation in balloon-injured dog carotid artery.
N. Shiota (1999)
Kinetics of association of serine proteinases with native and oxidized alpha-1-proteinase inhibitor and alpha-1-antichymotrypsin.
K. Beatty (1980)
Tryptase haplotype in mastocytosis: relationship to disease variant and diagnostic utility of total tryptase levels.
C. Akin (2007)
Kinetics of association of human proteinases with human alpha 2-macroglobulin.
G. D. Virca (1984)
Transgenic study of the function of chymase in heart remodeling
L. Chen (2002)
Highly efficient inhibition of human chymase by alpha(2)-macroglobulin.
M. Walter (1999)
Albumin Is a Substrate of Human Chymase
W. Raymond (2003)
The Significance of Chymase in the Progression of Abdominal Aortic Aneurysms in Dogs
K. Furubayashi (2007)
Tryptase genetics and anaphylaxis.
G. Caughey (2006)
Angiotensin I conversion by human and rat chymotryptic proteinases.
B. Wintroub (1984)
Impact of chymase inhibitor on cardiac function and survival after myocardial infarction.
D. Jin (2003)
Significance of Vascular Endothelial Cell Growth Factor Up-Regulation Mediated via a Chymase-Angiotensin-Dependent Pathway during Angiogenesis in Hamster Sponge Granulomas
J. Katada (2002)
Establishment of an assay method for human mast cell chymase.
K. Satomura (2002)
Tryptase and chymase: comparison of extraction and release in two dog mastocytoma lines.
G. Caughey (1988)
Angiotensin II induces hypertrophy of human airway smooth muscle cells: expression of transcription factors and transforming growth factor-beta1.
S. Mckay (1998)
Tryptase Precursors Are Preferentially and Spontaneously Released, Whereas Mature Tryptase Is Retained by HMC-1 Cells, Mono-Mac-6 Cells, and Human Skin-Derived Mast Cells 1
L. Schwartz (2003)
Mapping the extended substrate binding site of cathepsin G and human leukocyte elastase. Studies with peptide substrates related to the alpha 1-protease inhibitor reactive site.
K. Nakajima (1979)
Dipeptidyl Peptidase I Is Essential for Activation of Mast Cell Chymases, but Not Tryptases, in Mice*
P. Wolters (2001)
Angiotensin II generation by mast cell α- and β-chymases
G. Caughey (2000)
Usefulness of serum mast cell–specific chymase levels for postmortem diagnosis of anaphylaxis
H. Nishio (2005)
Kinetics of association of serine proteinases with native and oxidized 1 - proteinase inhibitor and 1 - antichymotryp
K. Beatty (1980)
The Three-dimensional Structure of the Human α2-Macroglobulin Dimer Reveals Its Structural Organization in the Tetrameric Native and Chymotrypsin α2-Macroglobulin Complexes*
S. Kolodziej (2002)
Angiotensin II induces hypertrophy of human airway smooth muscle cells: expression of transcription factors and transforming growth factor1
S. McKay (1998)
Significance of Chymase-Dependent Angiotensin II–Forming Pathway in the Development of Vascular Proliferation
M. Nishimoto (2001)
Compartmentalization of angiotensin II generation in the dog heart. Evidence for independent mechanisms in intravascular and interstitial spaces.
L. Dell'Italia (1997)
Mast cell chymase-like protease(s) modulates Escherichia coli lipopolysaccharide-induced vasomotor dysfunction in skeletal muscle in vivo.
H. Suzuki (1998)
Rapid conversion of angiotensin I to angiotensin II by neutrophil and mast cell proteinases.
C. Reilly (1982)
Human leukocyte cathepsin G. Subsite mapping with 4-nitroanilides, chemical modification, and effect of possible cofactors.
T. Tanaka (1985)
Angiotensin II-Forming Activity in a Reconstructed Ancestral Chymase
U. Chandrasekharan (1996)
Reaction of human skin chymotrypsin-like proteinase chymase with plasma proteinase inhibitors.
N. Schechter (1989)
Chymase as a proangiogenic factor: a possible involvement of chymase-angiotensin-dependent pathway in the hamster sponge angiogenesis model
M. Muramatsu (2000)
Kinetics of association of serine proteinases with native and oxidized 1-proteinase inhibitor and 1-antichymotrypsin
K. Beatty (1980)
The disappearance of enzyme-inhibitor complexes from the circulation of man.
K. Ohlsson (1976)
Involvement of chymase-mediated angiotensin II generation in blood pressure regulation.
Ming Li (2004)
Conditional and targeted overexpression of vascular chymase causes hypertension in transgenic mice
H. Ju (2001)
Involvement of chymasemediated angiotensin II generation in blood pressure regulation
M. Li (2004)
This paper is referenced by
Identification of a new alpha-2-macroglobulin: Multi-spectroscopic and isothermal titration calorimetry study.
A. Rehman (2016)
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A. Rehman (2013)
Overview of Mast Cells in Human Biology
Dean D Metcalfe (2020)
Protease Mediators of Anaphylaxis
G. Caughey (2011)
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S. Heuston (2012)
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S. Ahmad (2014)
Therapeutic targeting of cathepsin C: from pathophysiology to treatment
B. Korkmaz (2018)
Impact of Mast Cell Chymase on Renal Disease Progression.
H. Wasse (2012)
Biomarkers of the involvement of mast cells, basophils and eosinophils in asthma and allergic diseases
D. Metcalfe (2016)
Conformational behavior of alpha-2-macroglobulin: Aggregation and inhibition induced by TFE.
A. Rehman (2017)
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C. Craik (2010)
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Increased Plasma Chymase Concentration and Mast Cell Chymase Expression in Venous Neointimal Lesions of Patients with CKD and ESRD
H. Wasse (2011)
Mutational Tail Loss Is an Evolutionary Mechanism for Liberating Marapsins and Other Type I Serine Proteases from Transmembrane Anchors*
K. Raman (2013)
The αvβ6 integrin modulates airway hyperresponsiveness in mice by regulating intraepithelial mast cells.
K. Sugimoto (2012)
Mast cell peptidases: chameleons of innate immunity and host defense.
N. Trivedi (2010)
Haematopoietic Serine Proteases : A Cleavage Specificity Analysis
Michael Thorpe (2014)
Untersuchung der proteolytischen Prozessierung von humanen Angiotensin-Peptiden
D. Hildebrand (2013)
Mast cell proteases as pharmacological targets.
G. Caughey (2016)
Alpha‐2 macroglobulin as a region‐specific secretory protein in male reproductive tract, and its dynamics during sperm transit toward the female spermatheca in the blue crab
Thanyaporn Senarai (2017)
Specific Light-Up Probe with Aggregation-Induced Emission for Facile Detection of Chymase.
Ruoyu Zhang (2016)
α2-Macroglobulins: Structure and Function.
I. Garcia-Ferrer (2017)
Effects of angiotensin-converting enzyme inhibitor therapy on the regulation of plasma and cardiac tissue renin-angiotensin system in heart transplant patientsGlobal RAS regulation in HTx
J. J. (2016)
Biomarkers for evaluation of mast cell and basophil activation
K. Kabashima (2018)
Chapter 590 – Chymases
G. Caughey (2013)
Atheroinflammatory Properties of LDL and HDL Particles Modified by Human Mast Cell Neutral Proteases
Katariina Maaninka (2018)
Regulation of the pleiotropic effects of tissue-resident mast cells.
M. Huber (2019)
IgE-Mediated Systemic Anaphylaxis And Its Association With Gene Polymorphisms Of ACE, Angiotensinogen And Chymase
V. Varney (2019)
Biophysical analysis of interaction between curcumin and alpha-2-macroglobulin.
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