| Title: |
Prediction of a New Ligand-Binding Site for Type |
| Authors: |
Takeshi Takahashi; Hironori Suzuki; Tatsutoshi Inuzuka; Hideki Shibata; Masatoshi Maki |
| Contributors: |
The Pennsylvania State University CiteSeerX Archives |
| Source: |
http://www.mdpi.com/1422-0067/13/6/7532/pdf/. |
| Publication Year: |
2012 |
| Collection: |
CiteSeerX |
| Description: |
ALG-2 is a penta-EF-hand Ca 2+-binding protein and interacts with a variety of intracellular proteins. Two types of ALG-2-binding motifs have been determined: type 1, PXYPXnYP (X, variable; n = 4), in ALIX and PLSCR3; type 2, PXPGF, in Sec31A and PLSCR3. The previously solved X-ray crystal structure of the complex between ALG-2 and an ALIX peptide containing type 1 motif showed that the peptide binds to Pocket 1 and Pocket 2. Co-crystallization of ALG-2 and type 2 motif-containing peptides has not been successful. To gain insights into the molecular basis of type 2 motif recognition, we searched for a new hydrophobic cavity by computational algorithms using MetaPocket 2.0 based on 3D structures of ALG-2. The predicted hydrophobic pocket designated Pocket 3 fits with N-acetyl-ProAlaProGlyPhe-amide, a virtual penta-peptide derived from one of the two types of ALG-2-binding sites in PLSCR3 (type 2 motif), using the molecular docking software AutoDock Vina. We investigated effects of amino acid substitutions of the predicted binding sites on binding abilities by pulldown assays using glutathione-S-transferase-fused ALG-2 of wild-type and mutant proteins and lysates of cells expressing green fluorescent protein-fused PLSCR3 of wild-type and mutants. |
| Document Type: |
text |
| Language: |
English |
| Relation: |
http://citeseerx.ist.psu.edu/viewdoc/summary?doi=10.1.1.412.1692 |
| Availability: |
http://citeseerx.ist.psu.edu/viewdoc/summary?doi=10.1.1.412.1692; http://www.mdpi.com/1422-0067/13/6/7532/pdf/ |
| Rights: |
Metadata may be used without restrictions as long as the oai identifier remains attached to it. |
| Accession Number: |
edsbas.434C8D11 |
| Database: |
BASE |