Mast cells produce novel shorter forms of perlecan that contain functional endorepellin: a role in angiogenesis and wound healing

• Published in: The Journal of biological chemistry Vol. 288; no. 5; pp. 3289 - 3304
• Main Authors: Jung, Moonsun, Lord, Megan S, Cheng, Bill, Lyons, J Guy, Alkhouri, Hatem, Hughes, J Margaret, McCarthy, Simon J, Iozzo, Renato V, Whitelock, John M
• Format: Journal Article
• Language: English
• Published: United States American Society for Biochemistry and Molecular Biology 01.02.2013
• Subjects: Amino Acid Sequence; Animals; Cell Adhesion; Cell Line; Cell Movement; Coronary Vessels - cytology; Endothelial Cells - cytology; Endothelial Cells - metabolism; Glycobiology and Extracellular Matrices; Glycosaminoglycans - biosynthesis; Heparan Sulfate Proteoglycans - chemistry; Heparan Sulfate Proteoglycans - genetics; Heparan Sulfate Proteoglycans - isolation & purification; Heparan Sulfate Proteoglycans - metabolism; Humans; Integrin alpha2beta1 - metabolism; Lung - cytology; Mast Cells - cytology; Mast Cells - metabolism; Molecular Sequence Data; Neovascularization, Physiologic; Peptide Fragments - chemistry; Peptide Fragments - metabolism; Protein Isoforms - genetics; Protein Isoforms - metabolism; Protein Structure, Tertiary; Proteoglycans - biosynthesis; Rats; RNA, Messenger - genetics; RNA, Messenger - metabolism; Vesicular Transport Proteins - biosynthesis; Wound Healing
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M112.387811

Mast cells are derived from hematopoietic progenitors that are known to migrate to and reside within connective and mucosal tissues, where they differentiate and respond to various stimuli by releasing pro-inflammatory mediators, including histamine, growth factors, and proteases. This study demonstrated that primary human mast cells as well as the rat and human mast cell lines, RBL-2H3 and HMC-1, produce the heparan sulfate proteoglycan, perlecan, with a molecular mass of 640 kDa as well as smaller molecular mass species of 300 and 130 kDa. Utilizing domain-specific antibodies coupled with N-terminal sequencing, it was confirmed that both forms contained the C-terminal module of the protein core known as endorepellin, which were generated by mast cell-derived proteases. Domain-specific RT-PCR experiments demonstrated that transcripts corresponding to domains I and V, including endorepellin, were present; however, mRNA transcripts corresponding to regions of domain III were not present, suggesting that these cells were capable of producing spliced forms of the protein core. Fractions from mast cell cultures that were enriched for these fragments were shown to bind endothelial cells via the α(2)β(1) integrin and stimulate the migration of cells in "scratch assays," both activities of which were inhibited by incubation with either anti-endorepellin or anti-perlecan antibodies. This study shows for the first time that mast cells secrete and process the extracellular proteoglycan perlecan into fragments containing the endorepellin C-terminal region that regulate angiogenesis and matrix turnover, which are both key events in wound healing.