Effect of differentiation on the leucine enkephalin-degrading soluble enzymes released by the K562(S) cell line.
Leu-enkephalin hydrolysis kinetics were measured in the presence of soluble supernatants obtained from cultures of the K562(S) leukaemic cell line. Under these conditions, the substrate is degraded with formation of two distinct patterns of the hydrolysis by-products: in one pattern, similar amounts of Tyr and Tyr-Gly are formed; in the other, only Tyr-Gly can be measured. Kinetic data suggest that soluble proteolyses are released by these cells, and that either dipeptidylaminopeptidases alone, or both aminopeptidases and dipeptidylaminopeptidases are involved in substrate hydrolysis. This alternation of hydrolysis patterns appears consistent with existing data on the heterogeneity of K562 cells. In contrast with these results, chromatographic separation of the soluble enzymes indicates the release of all three classes of proteolyses known to hydrolyze enkephalins: aminopeptidases, dipeptidylaminopeptidases and dipeptidylcarboxypeptidases. In cells induced to differentiate by treatment with butyric acid, substrate hydrolysis is increased, and the pattern of the enzymes released is modified. In these cells, variations in both total proteolytic activity, and ratio between the three enzyme classes mentioned above are only minor, while the ratio between the different enzyme species within each class is greatly modified. Data obtained suggest that the expression of soluble enzymes is modified by differentiation. These data may also be interpreted as stressing the role of competition in controlling substrate hydrolysis by the multiple enzymes co-released by K562(S) cells.
Marini
M
M
Dept. Medicina Sperimentale, Universita' Tor Vergata, Roma, Italy.
Bongiorno
L
L
Urbani
A
A
Trani
E
E
Roda
L G
LG
D002454:Cell Differentiation; D005591:Chemical Fractionation; D004743:Enkephalin, Leucine / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000201:enzymology* / Q000378:metabolism / Q000473:pathology*; D012995:Solubility; D014407:Tumor Cells, Cultured
D004743:Enkephalin, Leucine