+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Determination of the primary structures of human skin chymase and cathepsin G from cutaneous mast cells of urticaria pigmentosa lesions



Determination of the primary structures of human skin chymase and cathepsin G from cutaneous mast cells of urticaria pigmentosa lesions



Journal of Immunology 152(8): 4062-4069



This study establishes the primary structure of human skin chymase and provides further evidence for the presence of a cathepsin G-like proteinase within human mast cells. The amino acid sequence of human skin chymase was established by protein methods and by analysis of PCR amplification products obtained with cDNA-derived from urticaria pigmentosa (UP) lesions. UP is a disease characterized by skin lesions containing high numbers of mast cells. Proteolytic digests of human chymase purified from normal skin yielded 10 resolvable peptides that were sequenced by automated Edman degradation. The amino acid sequences for these peptides combined with the sequence obtained for the protein's NH-2-terminal region (35 residues) accounted for 137 residues of the human skin chymase sequence. This partial amino acid sequence corresponded to the sequence of human heart chymase, a proteinase isolated from heart tissue with immunologic and hydrolytic properties similar to skin chymase. PCR amplification of UP-derived cDNA with primers based on the cDNA structure of heart chymase demonstrated a single amplification product of expected size which was subcloned and sequenced. The amino acid sequence (135 residues) deduced from this product was identical to that of heart chymase in the region between the primers. This sequence, along with that established for the purified protein, constituted 99% of the heart chymase primary structure, strongly indicating that human skin and heart chymases have identical primary structures. Amplification of the same UP-cDNA with primers coding for the NH-2- and COOH-terminal sequences of human neutrophil cathepsin G also produced a specific amplification product which was sequenced. The deduced amino acid sequence between the primers was identical to that reported for neutrophil cathepsin G, indicating that the protein of cutaneous mast cells previously shown to be immunologically cross-reactive with neutrophil cathepsin G has a comparable amino acid sequence. UP-cDNA demonstrating amplification products for cathepsin G did not demonstrate amplification products for human neutrophil elastase, suggesting that the cathepsin G PCR amplification product was not derived from neutrophils or monocytes possibly contaminating the lesion. These studies provide further evidence that human skin mast cells contain two different chymotrypsin-like proteinases.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 008448799

Download citation: RISBibTeXText

PMID: 8144971


Related references

The remarkable tissue mast cells; with observations on mast cell acid polysaccharides in the cutaneous lesions of urticaria pigmentosa. Archives of Dermatology 80: 725-730, 1959

Immunohistochemical characterization of human cutaneous mast cells in urticaria pigmentosa (cutaneous mastocytosis). Acta Pathologica Japonica 41(5): 344-349, 1991

C5a receptors are detectable on mast cells in normal human skin and in psoriatic plaques but not in weal and flare reactions or in urticaria pigmentosa by immunohistochemistry. Archives Of Dermatological Research. 289(2): 83-86, 1997

Remarkable cytoplasmic structures in mast cells of urticaria pigmentosa. Acta Dermato-Venereologica 50(1): 3-8, 1970

Electronmicroscopic Observations of Mast Cells in the Lesions of Urticaria Pigmentosa. Hifuka Kiyo. Acta Dermatologica 58: 132-135, 1963

Levels of tryptase, chymase, and Fc epsilon RI alpha messenger RNA in human skin are unchanged after IgE-dependent stimulation of cutaneous mast cells in vivo. Journal of Allergy and Clinical Immunology 99(2): 224-226, 1997

In vitro reactivity of mast cells in urticaria pigmentosa skin. Archives Of Dermatological Research. 290(1-2): 14-17,.-., 1998

The fine structure of skin mast cells in urticaria pigmentosa. Japanese Journal of Dermatology 79(2): 134-141, 1969

Direct histochemical demonstration of histamine in cutaneous mast cells: urticaria pigmentosa and keloids. Experientia 25(8): 854-855, 1969

Histamine release from skin mast cells and basophils in patients with urticaria pigmentosa. Acta Dermato-Venereologica 70(2): 154-156, 1990

Response of cutaneous mast cells to PUVA in patients with urticaria pigmentosa: histomorphometric, ultrastructural, and biochemical investigations. Journal of Investigative Dermatology 83(3): 175-178, 1984

CD25 expression on cutaneous mast cells from adult patients presenting with urticaria pigmentosa is predictive of systemic mastocytosis. American Journal of Surgical Pathology 32(1): 139-145, 2007

Retinoic acid inhibits in vitro development of mast cells but has no marked effect on mature human skin tryptase- and chymase-positive mast cells. Journal of Investigative Dermatology 120(2): 239-245, 2003

Inhibitors of chymase as mast cell-stabilizing agents: contribution of chymase in the activation of human mast cells. Journal of Pharmacology and Experimental Therapeutics 291(2): 517-523, 1999

The inhibition of mast cell activation by neutrophil lactoferrin: uptake by mast cells and interaction with tryptase, chymase and cathepsin G. Biochemical Pharmacology 65(6): 1007-1015, 2003