Journal of Otolaryngology and Ophthalmology of Shandong University ›› 2019, Vol. 33 ›› Issue (5): 40-43.doi: 10.6040/j.issn.1673-3770.0.2019.063

• Original Article • Previous Articles     Next Articles

Preliminary study on screening cochlear proteins interacting with myosin VI

Qingxiu YAO,Hui WANG,Zhuangzhuang LI,Dongzhen YU(),Shankai YIN   

  1. Department of Otolaryngology Head and Neck Surgery, Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, Shanghai 200233，China
  • Received:2019-02-15 Revised:2019-04-16 Online:2019-09-20 Published:2019-10-15
  • Contact: Dongzhen YU E-mail:drdzyu@126.com

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Abstract: Objective

To screen for cochlear proteins that interact with myosin VI.

Method

Cochlear proteins from SD rats were extracted for co-immunoprecipitation and mass spectrometry to screen for proteins interacting with myosin VI, which were further verified by immunofluorescence of whole mount basilar membrane and cryostat sections of cochleae.

 Result

Combined immunoprecipitation and mass spectrometry yielded more than 20 proteins interacting with myosin VI, and immunofluorescence showed that spectrin αII is primarily located at the cuticular plate and the lateral wall while myosin VI is mainly located in the cell body and cuticular plate. Both myosin VI and spectrin αII were co-localized in the cuticular plate.

 Conclusion

More than 20 cochlear proteins could interact with myosin VI, suggesting that myosin VI may interact with a variety of proteins to maintain stereociliary stability.

Key words: Spectrin αII, Myosin VI, Actin, Cuticular plate

CLC Number: 

  • R764.43

Fig. 1

SDS-PAGE silver stained strip after co-immunoprecipitation"

Table 1

The results of mass spectrometry"

编号 蛋白名称 蛋白质量 肽数 GI数
1 肌球蛋白-9样蛋白 227526.00 40 392349606
血影蛋白β链 274326.94 7 61557085
2 血影蛋白α链 285216.69 4 31543764
肌动蛋白 42051.85 4 4501885
3 热休克蛋白 83572.27 7 91234898
肌动蛋白 42051.85 4 4501885
4 血影蛋白α链 285216.69 4 31543764
肌动蛋白 42051.85 4 4501885
5 血影蛋白α链 285216.69 4 31543764
肌动蛋白 42051.85 4 4501885
6 微管蛋白α-1B链 50803.87 6 34740335
肌动蛋白 42051.85 4 4501885
7 微管蛋白α-1B链 50803.87 6 34740335
微管蛋白β-5链 50095.16 5 7106439
8 微管蛋白α-1B链 50803.87 6 34740335
血影蛋白α链 285216.69 4 31543764

Fig.2

The localization of myosin VI and spectrin αII"

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