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Digestion of C1q collagen‐like domain with MMPs‐1, ‐2, ‐3, and ‐9 further defines the sequence involved in the stimulation of neutrophil superoxide production
Author(s) -
Ruiz S.,
HenschenEdman A. H.,
Nagase H.,
Tenner A. J.
Publication year - 1999
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.66.3.416
Subject(s) - matrix metalloproteinase , superoxide , elastase , biology , trypsin , biochemistry , cleavage (geology) , neutrophil elastase , peptide sequence , microbiology and biotechnology , enzyme , inflammation , immunology , gene , paleontology , fracture (geology)
C1q, a subunit of the first component (C1) of the classical complement pathway, binds to neutrophils via its collagen‐like region (C1q‐CLR) stimulating superoxide production. We previously identified a region of C1q‐CLR, defined by fragments generated by trypsin and endoLys‐C digestion, that was required for triggering this respiratory burst. To further localize that critical site, purified human C1q was digested with pepsin to generate C1q‐CLR, and subsequently cleaved with the matrix metalloproteinases, MMP‐1, MMP‐2, MMP‐3, and MMP‐9. Digestion of C1q‐CLR with any of these MMPs did not alter the circular dichroism spectra, demonstrating that the fragments generated had maintained the secondary structure observed in the native molecule. All fragments retained the ability to trigger superoxide production by neutrophils. Analysis of the amino acid sequences of the purified cleavage products (none of which are identical to the published cleavage site specificities for these enzymes) demonstrated that it is the C‐chain, but not the A‐chain of C1q, that is critical for stimulating this activity, and thus may be a target for future therapeutic intervention. J. Leukoc. Biol. 66: 416–422; 1999.