caa a22 4500 397558376 CHVBK 20180308164809.0 cr unu---uuuuu 161202e199603 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021280 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021280 Differential Inhibition of Calpain and Proteasome Activities by Peptidyl Aldehydes of Di-Leucine and Tri-Leucine [Elektronische Daten] [Satoshi Tsubuki, Yumiko Saito, Masanori Tomioka, Hisashi Ito, Seiichi Kawashima] To explore membrane-permeable synthetic inhibitors that discriminate between endogenous calpain and proteasome in cells, we examined the inhibition profiles against calpain and proteasome in vitro and in vivo of peptidyl aldehydes possessing di-leucine and tri-leucine. The tripeptide aldehyde benzyloxycarbonyl-leucyl-leucyl-leucinal (ZLLLal) strongly inhibited calpain and proteasome activities in vitro. The concentration required for 50% inhibition (IC50) of the casein-degrading activity of calpain was 1.25 üM, and the IC50s for the succinyl-leucyl-leucyl-valyl-tyrosine-4-methylcoumaryl-7-amide (Suc-LLVY-MCA)- and benzyloxycarbonyl-leucyl-leucyl-leucine-4-methylcoumaryl-7-amide (ZLLL-MCA)-degrading activities of proteasome were 850 and 100 nM, respectively. On the . other hand, the synthetic dipeptide aldehyde benzyloxycarbonyl-leucyl-leucinal (ZLLal) strongly inhibited the casein degrading activity of calpain (IC501.20 μM), but the inhibition of proteasome was weak (ICsos for SucLLVY-MCA- and ZLLL-MCA-degrading activities were 120 and 110 μM, respectively). Thus, while calpain was inhibited by similar concentrations of ZLLal and ZLLLal, the inhibitory potencies of ZLLLal against the ZLLL-MCA-and SucLLVY-MCA-degrading activities in proteasome were 1,100 and 140 times stronger than those of ZLLal, respectively. To evaluate the effectiveness of these inhibitors on intracellular proteasome, the induction of neurite outgrowth in PC12 cells caused by proteasome inhibition was examined. ZLLLal and ZLLal initiated neurite outgrowth with optimal concentrations of 20 nM and 10 μM, respectively, again showing a big difference in the effective concentrations for the proteasome inhibition as in vitro. As for the effect on intracellular calpain, the concentrations of ZLLLal and ZLLal required for the inhibition of the autolytic activation of calpain in rabbit erythrocytes were 100 and 100 μM or more, respectively. The almost equal inhibitory potencies of ZLLLal and ZLLal were in agreement with the inhibition of calpain in vitro. These differential effects of inhibitors against calpain and proteasome are potentially useful for identifying the functions of calpain and proteasome in cell physiology and pathology © by the Journal of Biochemistry Regular Papers nationallicence calpain nationallicence neurite outgrowth nationallicence peptide aldehyde nationallicence protease inhibitor nationallicence proteasome nationallicence Tsubuki Satoshi Department of Molecular Biology, The Tokyo Metropolitan Institute of Medical Science (Rinsho-ken) 3-18-22 Honkomagome, Bunkyo-ku, Tokyo 113 aut Saito Yumiko Department of Molecular Biology, The Tokyo Metropolitan Institute of Medical Science (Rinsho-ken) 3-18-22 Honkomagome, Bunkyo-ku, Tokyo 113 aut Tomioka Masanori Department of Chemistry, Faculty of Science and Engineering, Aoyama Gakuin University 6-16-1 Chitosedai, Setagaya-ku, Tokyo 157 aut Ito Hisashi Department of Chemistry, Faculty of Science and Engineering, Aoyama Gakuin University 6-16-1 Chitosedai, Setagaya-ku, Tokyo 157 aut Kawashima Seiichi Department of Molecular Biology, The Tokyo Metropolitan Institute of Medical Science (Rinsho-ken) 3-18-22 Honkomagome, Bunkyo-ku, Tokyo 113 aut The Journal of Biochemistry Oxford University Press 119/3(1996-03), 572-576 0021-924X 119:3<572 1996 119 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021280 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021280 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Tsubuki Satoshi Department of Molecular Biology, The Tokyo Metropolitan Institute of Medical Science (Rinsho-ken) 3-18-22 Honkomagome, Bunkyo-ku, Tokyo 113 aut NATIONALLICENCE 700 1- Saito Yumiko Department of Molecular Biology, The Tokyo Metropolitan Institute of Medical Science (Rinsho-ken) 3-18-22 Honkomagome, Bunkyo-ku, Tokyo 113 aut NATIONALLICENCE 700 1- Tomioka Masanori Department of Chemistry, Faculty of Science and Engineering, Aoyama Gakuin University 6-16-1 Chitosedai, Setagaya-ku, Tokyo 157 aut NATIONALLICENCE 700 1- Ito Hisashi Department of Chemistry, Faculty of Science and Engineering, Aoyama Gakuin University 6-16-1 Chitosedai, Setagaya-ku, Tokyo 157 aut NATIONALLICENCE 700 1- Kawashima Seiichi Department of Molecular Biology, The Tokyo Metropolitan Institute of Medical Science (Rinsho-ken) 3-18-22 Honkomagome, Bunkyo-ku, Tokyo 113 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 119/3(1996-03), 572-576 0021-924X 119:3<572 1996 119 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford