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JKMRS Volume 25, No 2, pp 17, Backbone assignment of HMGB1 A-b...
2021년 06월 20일 / 조회수: 69

Backbone assignment of HMGB1 A-box and molecular interaction with Hoxc9DBD studied by paramagnetic probe

 

 

Ji Woong Choi and Sung Jean Park*

 

 

College of Pharmacy and Gachon Institute of Pharmaceutical Sciences, Gachon University, 191 Hambakmo-ero, Yeonsu-gu, Incheon 21936, Korea

 

  

Received May 28, 2021; Revised June 7, 2021; Accepted June 7, 2021

 

Abstract High mobility group protein B1 (HMGB1) is a highly conserved, non-histone, chromatin associated nuclear protein encoded by HMGB1 gene. HMGB1 proteins may be general co-factors in Hox-mediated transcriptional activation that facilitate the access of Hox proteins to specific DNA targets. It is unclear that the exact binding interface of Hoxc9DBD and HMGB1. To identify the interface and binding affinity of Hoxc9DBD and HMGB1 A-box, the paramagnetic probe, MTSL was used in NMR titration experiment. It is attached to the N-terminal end of HMGB1 A-box by reaction with thiol groups. The backbone assignment of HMGB1 A-box was achieved with 3D NMR techinques. The 15N-labeled HMGB1 A-box was titrated with MTSL-labeled Hoxc9DBD respectively. Based on the chemical shift changes we can identify the interacting residues and further map out the binding sites on the protein structure. The NMR titration result showed that the binding interface of HMGB1 A-box is around loop-1 between helix-1 and helix-2. In addition, the additional contacts were found in N- and C-terminus. The N-terminal arm region of Hoxc9DBD is the major binding region and the loop between helix1 and helix2 is the minor binding region.

 

Keywords Backbone assignment, HMGB1, Hoxc9, Paramagnetic probe 

 

 

* Address correspondence to: Sung Jean Park, College of Pharmacy and Gachon Institute of Pharmaceutical Sciences, Gachon University, 191 Hambakmo-ero, Yeonsu-gu, Incheon 21936, Korea Tel: 82-32-820-4957; E-mail: psjnmr@gachon.ac.kr 

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