Involvement of proteases in the action of IFN-gamma on WISH cells.
This study investigates the possible involvement of serine proteases in interferon-gamma (IFN-gamma) activity on WISH cells. It was observed that inhibition of (3)H-thymidine incorporation induced by IFN-gamma was abrogated by the serine protease inhibitors Nalpha-tosyl-L-lysyl-chloromethane and soybean trypsin inhibitor, both of which act mainly on trypsin. Phenylmethyl sulfonyl fluoride also had a partial inhibitory effect. Other protease inhibitors specific to the cysteine, the aspartic, and the metalloprotease families were not effective. Kinetic analysis revealed that a trypsin-like protease is involved in IFN-gamma activity for up to 7 h. Trypsin-like activity induced by IFN-gamma was detected in the particulate fraction but not in the cytosolic fraction, whereas chymotrypsin activity was not enhanced in either the cytosolic or particulate fractions under similar conditions. Following separation on a gelatin substrate gel, two trypsin-like protease activities located in the particulate fraction were found to increase in response to IFN-gamma treatment. Hence, it seems that a specific membrane-associated trypsin-like protease activity induced by IFN-gamma may play a role in the action of the cytokine on thymidine incorporation in WISH cells.
Aharon
Miri
M
Department of Clinical Biochemistry, Faculty of Health Sciences, Ben-Gurion University and Institute of Hematology, Soroka University Medical Center, Beer Sheva, Israel.
Dvilansky
Alexander
A
Shpilberg
Ofer
O
Nathan
Ilana
I
D000650:Amnion / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line / Q000187:drug effects; D002462:Cell Membrane / Q000201:enzymology; D004261:DNA Replication / Q000187:drug effects; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology*; D007976:Leupeptins / Q000494:pharmacology; D008239:Lysine / Q000031:analogs & derivatives* / Q000494:pharmacology; D010436:Pepstatins / Q000494:pharmacology; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000494:pharmacology; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000502:physiology*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D013347:Subcellular Fractions / Q000201:enzymology; D015244:Thiorphan / Q000494:pharmacology; D014107:Tosyllysine Chloromethyl Ketone / Q000494:pharmacology; D014358:Trypsin Inhibitor, Bowman-Birk Soybean / Q000494:pharmacology
D007976:Leupeptins; D010436:Pepstatins; D011480:Protease Inhibitors; D015842:Serine Proteinase Inhibitors; D014358:Trypsin Inhibitor, Bowman-Birk Soybean; C013791:N-acetylphenylalanine beta-naphthyl ester; D014107:Tosyllysine Chloromethyl Ketone; D010649:Phenylalanine; D010664:Phenylmethylsulfonyl Fluoride; C002306:N-tosyllysine methyl ester; D007371:Interferon-gamma; D015244:Thiorphan; D012697:Serine Endopeptidases; C032854:leupeptin; D008239:Lysine; C031375:pepstatin