Demonstration of Concanavalin A Receptors on Leptomonas pessoai Cell Membrane*§

SYNOPSIS. Leptomonas pessoai promastigotes cultivated in a synthetic medium were agglutinated with concanavalin A (Con A). Agglutination was predominantly of the flagellar-flagellar type, and was inhibited with sucrose or α-methyl-D-mannoside. Cell surface polysaccharides and/or glycoproteins were demonstrated by several cytochemical methods at the fine-structural level. A Con A-horseradish peroxidase-diaminobenzidine (DAB) technic was used to detect Con A receptors in the pellicular membrane of the flagellar pocket region and in the flagellar membrane proper. Somewhat less of the Con A-DAB reaction product was observed in the pellicular membrane enclosing the rest of the cell.     

Trypanosomatidae Isolated from Zelus leucogrammus (Perty, 1834) (Hemiptera,Reduviidae), with a Discussion on Flagellates of Insectivorous Bugs*

The insectivorous bug Zelus leucogrammus (Perty, 1834) (Hemiptera, Reduviidae) was found to harbor several species of trypanosomatids of the genera Leptomonas, Crithidia, Herpetomonas and Blastocrithidia. However, laboratory bred nymphs given sterile food were infected only by Blastocrithidia, the other flagellates appearing in their intestinal tract when they were fed on other insects. The Blastocrithidia was thought to belong to a single species and to be a parasite proper of Z. leucogrammus (and possibly other Hemiptera), producing luxuriant infections in the intestinal tract of the bugs captured in nature and spreading easily among isolated populations through cannibalism and ingestion of cysts; it was also abundant in the genital tract of gravid females but transovarian transmission was not proved. This Blastocrithidia experimentally infected Triatoma infestans and is very close to B. triatomae Cerisola, del Prado, Rohwedder and Bozzini, 1971. The other trypanosomatids, never numerous in naturally infected Z. leucogrammus, are acquired by the bug from many different insects, chiefly Diptera, on which it was seen to feed. Herpetomonas muscarum and C. luciliae, the common trypanosomatids among several muscoid Diptera, were isolated both from flies and from the bugs fed on them.     

Degradation of protein kinase Cα and its free catalytic subunit, protein kinase M, in intact human neuroblastoma cells and under cell-free conditions: Evidence that PKM is degraded by mM calpain-mediated proteolysis at a faster rate than PKC

Proteolytic cleavage of protein kinase C (PKC) under cell-free conditions generates a co-factor independent, free catalytic subunit (PKM). However, the difficulty in visualizing PKM in intact cells has generated controversy regarding its physiological relevance. In the present study, treatment of SH-SY-5Y cells with 2-O-tetradecanoylphorbol 13-acetate resulted in complete down-regulation of PKC within 24 h without detection of PKM. By contrast, low levels of PKM were transiently detected following ionophore-mediated calcium influx under conditions which induced no detectable PKC loss. PKM was not detected during rapid cell-free degradation of partially purified SH-SY-5Y PKCα by purified human brain mM calpain. However, when the kinetics of PKC degradation were slowed by lowering levels of calpain, PKM was transiently detected. PKM was also only transiently observed following calpain-mediated degradation of purified rat brain PKCα. Densitometric analyses indicated that, once formed, PKM was degraded approximately 10 times faster than PKC. These data provide an explanation as to why PKM is difficult to observe in situ, and indicate that PKM should not be considered as an ‘unregulated’ kinase, since its persistence is apparently strictly regulated by proteolysis.