IN-VITRO RECONSTITUTION OF PHAGOSOME-ENDOSOME FUSION - EVIDENCE FOR REGULATION BY HETEROTRIMERIC GTPASES

被引:35
作者
BERON, W
COLOMBO, MI
MAYORGA, LS
STAHL, PD
机构
[1] WASHINGTON UNIV,SCH MED,DEPT CELL BIOL & PHYSIOL,ST LOUIS,MO 63110
[2] UNIV NACL CUYO,FAC CIENCIAS MED,CONICET,INST HISTOL & EMBRIOL,MENDOZA,ARGENTINA
关键词
HETEROTRIMERIC GTPASES; INTRACELLULAR TRANSPORT; PHAGOCYTOSIS;
D O I
10.1006/abbi.1995.1172
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
We have assessed the role of heterotrimeric GTPases on in vitro fusion of phagosomes and endosomes. Highly purified phagosomes were found to contain G alpha s, G alpha i1, G alpha i2, G alpha i3, and G beta subunits of heterotrimeric GTP-binding proteins. A functional role for G proteins was established using an in vitro phagosome-endosome fusion assay, First, addition of AlF4- and purified G beta gamma subunits to the in vitro assay blocked fusion, indicating that heterotrimeric G proteins may play a role, either direct or indirect, in phagosome maturation. Second, a striking inhibitory effect was observed when the vesicles were incubated with peptides that preferentially activate G alpha s. A similar effect on phagosome-endosome fusion was observed with cholera toxin, a reagent known to activate G alpha s. Our results suggest that one or more heterotrimeric G proteins, including Gs, mediate and/or regulate phagosome-endosome fusion. (C) 1995 Academic Press, Inc.
引用
收藏
页码:337 / 342
页数:6
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