title: Heparin is essential for the storage of specific granule proteases in mast cells.
abstract: All mammals produce heparin, a negatively charged glycosaminoglycan that is a major constituent of the secretory granules of mast cells which are found in the peritoneal cavity and most connective tissues. Although heparin is one of the most studied molecules in the body, its physiological function has yet to be determined. Here we describe transgenic mice, generated by disrupting the N-deacetylase/N-sulphotransferase-2 gene, that cannot express fully sulphated heparin. The mast cells in the skeletal muscle that normally contain heparin lacked metachromatic granules and failed to store appreciable amounts of mouse mast-cell protease (mMCP)-4, mMCP-5 and carboxypeptidase A (mMC-CPA), even though they contained substantial amounts of mMCP-7. We developed mast cells from the bone marrow of the transgenic mice. Although these cultured cells contained high levels of various protease transcripts and had substantial amounts of mMCP-6 protein in their granules, they also failed to express mMCP-5 and mMC-CPA. Our data show that heparin controls, through a post-translational mechanism, the levels of specific cassettes of positively charged proteases inside mast cells.
authors.affiliation: Department of Medicine, Boston University School of Medicine, and VA Medical Center, Massachusetts 02130, USA. Humphries.Donald_E@BOSTON.VA.GOV
authors.forename: D E
authors.initials: DE
authors.lastname: Humphries
authors.forename: G W
authors.initials: GW
authors.lastname: Wong
authors.forename: D S
authors.initials: DS
authors.lastname: Friend
authors.forename: M F
authors.initials: MF
authors.lastname: Gurish
authors.forename: W T
authors.initials: WT
authors.lastname: Qiu
authors.forename: C
authors.initials: C
authors.lastname: Huang
authors.forename: A H
authors.initials: AH
authors.lastname: Sharpe
authors.forename: R L
authors.initials: RL
authors.lastname: Stevens
mesh_terms: D016475:3T3 Cells; D000581:Amidohydrolases / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001854:Bone Marrow Cells / Q000201:enzymology; D002268:Carboxypeptidases / Q000378:metabolism; D043422:Carboxypeptidases A; D002454:Cell Differentiation; D002478:Cells, Cultured; D053818:Chymases; D018920:Coculture Techniques; D003594:Cytoplasmic Granules / Q000201:enzymology / Q000502:physiology*; D004792:Enzyme Precursors / Q000378:metabolism; D006493:Heparin / Q000502:physiology*; D007413:Intestinal Mucosa / Q000166:cytology; D007583:Jejunum / Q000166:cytology; D008407:Mast Cells / Q000166:cytology / Q000201:enzymology / Q000276:immunology / Q000502:physiology*; D051379:Mice; D008822:Mice, Transgenic; D012697:Serine Endopeptidases / Q000378:metabolism*; D013431:Sulfates / Q000378:metabolism; D015238:Sulfotransferases / Q000235:genetics / Q000378:metabolism; D017160:Trichinella spiralis; D014235:Trichinellosis / Q000276:immunology; D053802:Tryptases
chemical_list: D004792:Enzyme Precursors; D013431:Sulfates; C509190:Tpsb2 protein, mouse; D006493:Heparin; C491855:Ndst2 protein, mouse; D015238:Sulfotransferases; D002268:Carboxypeptidases; D043422:Carboxypeptidases A; D012697:Serine Endopeptidases; C066932:mast cell protease 4; D053818:Chymases; C509222:Tpsab1 protein, mouse; D053802:Tryptases; D000581:Amidohydrolases