標題: The First Residue of the PWWP Motif Modulates HATH Domain Binding, Stability, and Protein-Protein Interaction
作者: Hung, Yi-Lin
Lee, Hsia-Ju
Jiang, Ingjye
Lin, Shang-Chi
Lo, Wei-Cheng
Lin, Yi-Jan
Sue, Shih-Che
生物資訊及系統生物研究所
Institude of Bioinformatics and Systems Biology
公開日期: 7-七月-2015
摘要: Hepatoma-derived growth factor (hHDGF) and HDGF-related proteins (HRPs) contain conserved N-terminal HATH domains with a characteristic structural motif; namely the PWWP motif. The HATH domain has attracted attention because of its ability to bind with heparin/heparan sulfate, DNA, and methylated histone peptide. Depending on the sequence of the PWWP motif, HRP HATHs are classified into P-type (Pro-His-Trp-Pro) and A-type (Ala-His-Trp-Pro) forms. A-type HATH is highly unstable and tends to precipitate in solution. We replaced the Pro residue in P-type HATH(HDGF) with Ala and evaluated the influence on structure, dynamics, and ligand binding. Nuclear magnetic resonance (NMR) hydrogen/deuterium exchange and circular dichroism (CD) measurements revealed reduced stability. Analysis of NMR backbone N-15 relaxations (R-1, R-2, and nuclear Overhauser effect) revealed additional backbone dynamics in the interface between the beta-barrel and the C-terminal helix bundle. The beta 1-beta 2 loop, where the AHWP sequence is located, has great structural flexibility, which aids HATH-HATH interaction through the loop. A-type HATH, therefore, shows a stronger tendency to aggregate when binding with heparin and DNA oligomers. This study defines the role of the first residue of the PWWP motif in modulating HATH domain stability and oligomer formation in binding.
URI: http://dx.doi.org/10.1021/acs.biochem.5b00454
http://hdl.handle.net/11536/128043
ISSN: 0006-2960
DOI: 10.1021/acs.biochem.5b00454
期刊: BIOCHEMISTRY
Volume: 54
Issue: 26
起始頁: 4063
結束頁: 4074
顯示於類別:期刊論文