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Studies on glycyl-proline naphthylamidase

I. Lymphocytes

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Summary

Glycyl-proline naphthylamidase (Gly-Pro-Nase) was discovered in 39.4% (±3.4) of peripheral blood lymphocytes of healthy adult humans and in 38.5% (±3.1) of peripheral blood lymphocytes of mini-pigs using glycyl-L-proline-4-methoxy-2-naphthylamide as the substrate and Fast Blue B (diazonium salt to be preferred) or hexazotized New Fuchsin as the coupling agents. The pH optimum of the enzyme is in the alkaline range in the vicinity of neutral. The enzyme activity in lymphocytes is not influenced by 10−3 M EDTA, 10−3 M N-ethyl-maleimide, and 10−3 M MnCl2. It is inhibited to about 50% by 1.4·10−4 M Pb (NO3)2. Individual lymphocytes differ in their activity. In some lymphocytes only one small positive dot in the cytoplsm can be seen. In other cells the number of these dots is greater. In other cases the cytoplasm is overfilled with positively reacting granules and rods. Gly-Pro-Nase in lymphocytes is confined to lysosomes. These organelles may not be its exclusive localization, however.

The activity of Gly-Pro-Nase is present in lymphocytes forming E-rosettes with sheep erythrocytes. There is no correlation between the number of bound erythrocytes and the enzyme activity. An unequivocal presence of this enzyme in B-lymphocytes remains to be established. Gly-Pro-Nase is present in differentiated lymphocytes particularly in those bearing ring-shaped nucleoli. It was demonstrated neither in the blastically transformed lymphocytes (after phytohemagglutinine stimulation) nor in epithelial lymphocytes of the human jejunum. Its activity does not go parallel to that of acid esterase or acid phosphatase.

Preliminary investigation on peripheral blood lymphocytes of patients sufferring of various diseases of the hemopoetic system revealed a decreased number of positively reacting lymphocytes in chronic lymphatic leukemia (1–20%), normal or slightly elevated values in chronic myeloid leukemia (40–68%), highly elevated values in myelofibrosis (60–78%), and decreased, normal or elevated values in lymphogranuloma (0–70%).

Studies on the metabolic as well as diagnostic significance of Gly-Pro-Nase activity in lymphocytes are in progress.

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  1. Research Laboratory of Angiology, 1st Department of Pathology, Faculty of General Medicine, Charles University, Studničkova 2, 12800, Praha 2, ČSSR

    Zdeněk Lojda

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  1. Zdeněk Lojda
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Lojda, Z. Studies on glycyl-proline naphthylamidase. Histochemistry 54, 299–309 (1977). https://doi.org/10.1007/BF00508273

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