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Isolation and Characterization of Myeloperoxidase from Leukocytes of Rat Peritoneal Fluid

V. I. Morozov,1,2 P. V. Tsyplenkov,1 V. N. Kokryakov,3 K. N. Volkov,1 and N. A. Vinogradova4

1Department of Biochemistry, Institute of Physical Culture, pr. Dinamo 2, St. Petersburg, 197042 Russia; fax: (7-812) 235-01-31.

2To whom correspondence should be addressed.

3Department of General Pathology and Pathophysiology, Institute of Experimental Medicine, Russian Academy of Medical Sciences, ul. Akademika Pavlova 12, St. Petersburg, 197376 Russia; fax: (7-812) 234-94-93.

4Laboratory of Physiology of Cell Cycle, Institute of Cytology, Russian Academy of Sciences, Tikhoretskii pr. 4, St. Petersburg, 194064 Russia; fax: (7-812) 247-03-41.

Submitted September 12, 1996; revision submitted February 7, 1997.
Myeloperoxidase (MPO) was isolated from rat peritoneal leukocytes with a yield of 51% and A430/A280 = 0.75-0.80, and its physicochemical properties were studied. The molecular weight of the MPO is about 150 kD. The MPO was assayed for amino acid content. We used substrate mixture containing phenol, 4-aminoantipyrine, and H2O2 to detect 10-10 M of the enzyme. The MPO was localized in rat blood neutrophils using polyclonal anti-MPO antibodies and secondary fluorescein isothiocyanate-labeled antibodies. Immunofluorimetric assay (IFMA) was developed for quantitative measurement of the MPO. The MPO and leukocytes can iodinate BSA using NaI or thyroxine as the source of iodine.
KEY WORDS: leukocytes, neutrophils, myeloperoxidase, iodination.