Per-deuteration and NMR experiments for the backbone assignment of 62 kDa protein, Hsp31

Kim, Jihong;Choi, Dongwook;Park, Chankyu;Ryu, Kyoung-Seok

  • 투고 : 2015.10.07
  • 심사 : 2015.11.29
  • 발행 : 2015.12.20


Hsp31 protein is one of the members of DJ-1 superfamily proteins and has a dimeric structure of which molecular weight (MW) is 62 kDa. The mutation of DJ-1 is closely related to early onset of Parkinson's disease. Hsp31 displays $Zn^{+2}$-binding activity and was first reported to be a holding chaperone in E. coli. Its additional glyoxalase III active has recently been characterized. Moreover, an incubation at $60^{\circ}C$ induces Hsp31 protein to form a high MW oligomer (HMW) in vitro, which accomplishes an elevated holding chaperone activity. The NMR technique is elegant method to probe any local or global structural change of a protein in responses to environmental stresses (heat, pH, and metal). Although the presence of the backbone chemical shifts (bbCSs) is a prerequisite for detailed NMR analyses of the structural changes, general HSQC-based triple resonance experiments could not be used for 62 kDa Hsp31 protein. Here, we prepared the per-deuterated Hsp31 and performed the TROSY-based triple resonance experiments for the bbCSs assignment. Here, detailed processes of per-deuteration and the NMR experiments are described for other similar NMR approaches.


Backbone chemical shift assignment;Hsp31;NMR;Per-deuteration;TROSY


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피인용 문헌

  1. Backbone resonance assignments of the Escherichia coli 62 kDa protein, Hsp31 vol.11, pp.2, 2017,


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