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Guanine nucleotide exchange factor 2 for Rab5 proteins coordinated with GLUP6/GEF regulates the intracellular transport of the proglutelin from the Golgi apparatus to the protein storage vacuole in rice endosperm.

Wen L, Fukuda M, Sunada M, Ishino S, Ishino Y, Okita TW, Ogawa M, Ueda T, Kumamaru T - J. Exp. Bot. (2015)

Bottom Line: Like GLUP6/GEF, Rab5-GEF2 was capable of activating Rab5a and two other Rab5 isoforms in in vitro GTP/GDP exchange assays.GEF proteins consist of the helical bundle (HB) domain at the N-terminus, Vps9 domain, and a C-terminal region.By the deletion analysis of GEFs, the HB domain was found essential for the activation of Rab5 proteins.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan Tobacco Research Institute, Chinese Academy of Agricultural Science, Qingdao 266101, China.

No MeSH data available.


Related in: MedlinePlus

in vitro GEF assay of Rab5-GEF2. The conformational changes of Rab5s and Rab11 (as 1 μM GST fusion proteins) upon GDP/GMP-PNP exchange were measured by monitoring Trp autofluorescence when incubated with 0 μM (blue), 0.25 μM (magenta), 0.5μM (yellow), and 1 μM (red) GST-Rab5-GEF2. Note that GST-Rab5-GEF2 activates GLUP4/Rab5a, Rab5b, and Rab5c, but not Rab11.
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Figure 7: in vitro GEF assay of Rab5-GEF2. The conformational changes of Rab5s and Rab11 (as 1 μM GST fusion proteins) upon GDP/GMP-PNP exchange were measured by monitoring Trp autofluorescence when incubated with 0 μM (blue), 0.25 μM (magenta), 0.5μM (yellow), and 1 μM (red) GST-Rab5-GEF2. Note that GST-Rab5-GEF2 activates GLUP4/Rab5a, Rab5b, and Rab5c, but not Rab11.

Mentions: In order to elucidate whether the Rab5-GEF2 can activate Rab5s including GLUP4/Rab5a, in vitro guanine nucleotide exchange assays were conducted. As expected, Rab5-GEF2 activated GLUP4/Rab5a, the plant-specific Rab5b, and Rab5c in a dose-dependent manner, while the control Rab11 was not activated by Rab5-GEF2 (Fig. 7). This result indicates that Rab5-GEF2 serves as a general activator of Rab5 isoforms. These results provide ample evidence that both GLUP6/GEF and Rab5-GEF2 activate multiple Rab5 proteins present in developing rice endosperm.


Guanine nucleotide exchange factor 2 for Rab5 proteins coordinated with GLUP6/GEF regulates the intracellular transport of the proglutelin from the Golgi apparatus to the protein storage vacuole in rice endosperm.

Wen L, Fukuda M, Sunada M, Ishino S, Ishino Y, Okita TW, Ogawa M, Ueda T, Kumamaru T - J. Exp. Bot. (2015)

in vitro GEF assay of Rab5-GEF2. The conformational changes of Rab5s and Rab11 (as 1 μM GST fusion proteins) upon GDP/GMP-PNP exchange were measured by monitoring Trp autofluorescence when incubated with 0 μM (blue), 0.25 μM (magenta), 0.5μM (yellow), and 1 μM (red) GST-Rab5-GEF2. Note that GST-Rab5-GEF2 activates GLUP4/Rab5a, Rab5b, and Rab5c, but not Rab11.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4588877&req=5

Figure 7: in vitro GEF assay of Rab5-GEF2. The conformational changes of Rab5s and Rab11 (as 1 μM GST fusion proteins) upon GDP/GMP-PNP exchange were measured by monitoring Trp autofluorescence when incubated with 0 μM (blue), 0.25 μM (magenta), 0.5μM (yellow), and 1 μM (red) GST-Rab5-GEF2. Note that GST-Rab5-GEF2 activates GLUP4/Rab5a, Rab5b, and Rab5c, but not Rab11.
Mentions: In order to elucidate whether the Rab5-GEF2 can activate Rab5s including GLUP4/Rab5a, in vitro guanine nucleotide exchange assays were conducted. As expected, Rab5-GEF2 activated GLUP4/Rab5a, the plant-specific Rab5b, and Rab5c in a dose-dependent manner, while the control Rab11 was not activated by Rab5-GEF2 (Fig. 7). This result indicates that Rab5-GEF2 serves as a general activator of Rab5 isoforms. These results provide ample evidence that both GLUP6/GEF and Rab5-GEF2 activate multiple Rab5 proteins present in developing rice endosperm.

Bottom Line: Like GLUP6/GEF, Rab5-GEF2 was capable of activating Rab5a and two other Rab5 isoforms in in vitro GTP/GDP exchange assays.GEF proteins consist of the helical bundle (HB) domain at the N-terminus, Vps9 domain, and a C-terminal region.By the deletion analysis of GEFs, the HB domain was found essential for the activation of Rab5 proteins.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan Tobacco Research Institute, Chinese Academy of Agricultural Science, Qingdao 266101, China.

No MeSH data available.


Related in: MedlinePlus