ATP-DEPENDENT REVERSIBLE ASSOCIATION OF PROTEASOMES WITH MULTIPLE PROTEIN-COMPONENTS TO FORM 26S COMPLEXES THAT DEGRADE UBIQUITINATED PROTEINS IN HUMAN HL-60 CELLS

被引:149
作者
ORINO, E
TANAKA, K
TAMURA, T
SONE, S
OGURA, T
ICHIHARA, A
机构
[1] UNIV TOKUSHIMA,INST ENZYME RES,TOKUSHIMA 770,JAPAN
[2] UNIV TOKUSHIMA,SCH MED,DEPT INTERNAL MED 3,TOKUSHIMA 770,JAPAN
关键词
PROTEASOME; 26S PROTEASE COMPLEX; ATP; UBIQUITIN; HL-60; CELL;
D O I
10.1016/0014-5793(91)80686-W
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The role of proteasomes in ubiquitin (Ub)-dependent protein degradation was studied by analyzing lysates of human promyelocytic leukemia HL-60 cells by glycerol density gradient centrifugation. High succinyl-Leu-Leu-Val-Tyr-4-methylcoumaryl-7-amide hydrolyzing activity was found in the 26S fraction, whereas the 20S fraction containing proteasomes had no activity. Addition of 0.05% sodium dodecylsulfate to the latter fraction, however, induced marked activity. The 26S, but not the 20S fraction catalyzed ATP-dependent degradation of [I-125]lysozyme-Ub conjugate. Depletion from the lysate of ATP caused complete shift of the active 26S complex to the latent 20S form, whereas in the lysate prepared from ATP-depleted cells, ATP converted 20S proteasomes to 26S complexes. The immunoprecipitated 26S complexes were found to consist of proteasomes and 13 15 other proteins ranging in size from 35 to 110 kDa. We conclude that in the lysate, latent proteasomes undergo reversible, ATP-dependent association with multiple protein components to form 26S complexes that catalyze ATP-dependent degradation of Ub-protein conjugates.
引用
收藏
页码:206 / 210
页数:5
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