Introduction Directory UMM :Data Elmu:jurnal:P:PlantScience:PlantScience_Elsevier:Vol149.Issue1.2000:
Plant Science 149 1999 33 – 41
Purification and characterization of the 26S proteasome from cultured rice Oryza sati6a cells
Yuki Yanagawa
a,b
, Akiko Ohhashi
a
, Yasuko Murakami
c
, Yasushi Saeki
d
, Hideyoshi Yokosawa
d
, Keiji Tanaka
e
, Junji Hashimoto
b,f
, Takahide Sato
a
, Hiroki Nakagawa
a,
a
Department of Bioproduction Science, Faculty of Horticulture, Chiba Uni6ersity, Matsudo, Chiba
271
-
8510
, Japan
b
CREST Core Research for E6olutional Science and Technology
of Japan Science and Technology Corporation JST
, Chiyoda-ku, Tokyo
101
-
0062
, Japan
c
Department of Biochemistry
2
, The Jikei Uni6ersity School of Medicine, Nishishinbashi, Minato-ku, Tokyo
105
-
8461
, Japan
d
Department of Biochemistry, Graduate School of Pharmaceutical Sciences, Hokkaido Uni6ersity, Kita-ku, Sapporo
060
-
0812
, Japan
e
The Tokyo Metropolitan Institute of Medical Science, Honkomagome, Bunkyo-ku, Tokyo
113
-
8613
, Japan
f
National Institute of Agrobiological Resources, Kannondai, Tsukuba, Ibaraki
305
-
8602
, Japan Received 30 April 1999; received in revised form 12 July 1999; accepted 13 July 1999
Abstract
The 26S proteasome was purified from cultured rice cells to near homogeneity by ultracentrifugation for 5 h at 85,000 × g, chromatography on Biogel A-1.5m, and glycerol density-gradient centrifugation analysis. The purified enzyme had two distinct
forms, termed 26Sa- and 26Sb-type proteasomes, with different electrophoretic mobilities by nondenaturing polyacrylamide gel electrophoresis. It consisted of multiple polypeptides with molecular masses of 25 – 35 and 42 – 120 kDa, which presumably
corresponded to those of the 20S proteasome and an associated PA700 regulatory complex, respectively. The rice 26S proteasome resembled, but was not identical to, one from other sources in their subunit composition and immunochemical reactivity.
Intriguingly, both rice and spinach 26S proteasomes could not degrade rat ornithine decarboxylase in the presence of antizyme and ATP, unlike the rat 26S proteasome, implying the existence of functional differences between mammalian and plant 26S
proteasomes. © 1999 Published by Elsevier Science Ireland Ltd. All rights reserved.
Keywords
:
26S Proteasome; 20S Proteasome; Rice Oryza sati6a L. www.elsevier.comlocateplantsci