J. Biol. Chem.

Myeloperoxidase (MPO) is a functionally important component of the normal human neutrophil host defense system. This enzyme possesses a dimeric structure composed of two heavy subunit (55-63 kDa)/light subunit (10-15 kDa) protomers, each of which is associated with a heme-like prosthetic group. In addition, MPO species of approximately 38 and 22 kDa have been reported by many different investigators, but their nature and mode of origin are not understood. In the present study, we demonstrate that when MPO is heated under nonreducing, denaturing conditions, these two species are produced via a novel autolytic cleavage of the heavy subunit. The 38-kDa species was isolated by fast-protein liquid chromatography and identified by sequencing as the carboxyl-terminal portion of the heavy subunit, and the cleavage was shown to occur exclusively between Met409 and Pro410. In order to further characterize this unusual cleavage reaction, the 22-kDa species was digested with endoproteinase Asp-N, and the peptide corresponding to its carboxyl terminus was isolated and analyzed by sequencing and mass spectrometry. These data indicated that during cleavage of the heavy subunit, Met409 was converted to homoserine lactone. Thus, the cleavage appeared to formally resemble the cyanogen bromide-dependent cleavage of Met-X peptide bonds. Recent x-ray crystallographic data for canine MPO have indicated that Met409 is in close proximity to the heme-like prosthetic group of MPO. Our studies suggest that interaction of Met409 with this group leads to the formation of a methionyl sulfonium derivative which undergoes intramolecular rearrangement with subsequent peptide cleavage under nonreducing conditions. This derivative may be, at least in part, responsible for the unusual spectral characteristics and enzymatic properties of the enzyme. The primary structure of the 22-kDa MPO species is also reported, and direct evidence is provided for asparagine-linked oligosaccharide moieties at two of the three predicted glycosylation sites.

Source:http://purl.uniprot.org/citations/1334087