Abstract
Yeast Doa1/Ufd3 is an adaptor protein for Cdc48 (p97 in mammal), an AAA type ATPase associated with endoplasmic reticulum-associated protein degradation pathway and endosomal sorting into multivesicular bodies. Doa1 functions in the endosomal sorting by its association with Hse1, a component of endosomal sorting complex required for transport (ESCRT) system. The association of Doa1 with Hse1 was previously reported to be mediated between PFU domain of Doa1 and SH3 of Hse1. However, it remains unclear which residues are specifically involved in the interaction. Here we report that Doa1/PFU interacts with Hse1/SH3 with a moderate affinity of 5 μM. Asn-438 of Doa1/PFU and Trp-254 of Hse1/SH3 are found to be critical in the interaction while Phe-434, implicated in ubiquitin binding via a hydrophobic interaction, is not. Small-angle X-ray scattering measurements combined with molecular docking and biochemical analysis yield the solution structure of the Doa1/PFU:Hse1/SH3 complex. Taken together, our results suggest that hydrogen bonding is a major determinant in the interaction of Doa1/PFU with Hse1/SH3.
| Original language | English |
|---|---|
| Pages (from-to) | 352-357 |
| Number of pages | 6 |
| Journal | Biochemical and Biophysical Research Communications |
| Volume | 446 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 2014 Mar 28 |
Bibliographical note
Funding Information:We thank the staff at the beamline 4C of Pohang Accelerator Laboratory for technical assistance in collecting SAXS data. This work was supported by Basic Science Research Program ( 2009-0074586 and 2011-0023402 ) and the Pioneer Research Center Program ( 2012-0009597 ) through the National Research Foundation of Korea (NRF) and the Woo Jang Chun Program (PJ009106) through the Rural Development Agency.
All Science Journal Classification (ASJC) codes
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology