CRCU

Chemical Research in Chinese Universities ›› 2013, Vol. 29 ›› Issue (2): 280-284.doi: 10.1007/s40242-013-2329-y

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Purification of Complex of SAV1 with PP1A

XIN Hua1, ZHANG Yu2, LI Lin3, ZHANG Li-ping1   

  1. 1. Department of Thoracic Surgery, China-Japan Union Hospital of Jilin University, Changchun 130033, P. R. China;
    2. Department of Clinical Laboratory, the Second Hospital of Jilin University, Changchun 130041, P. R. China;
    3. Department of Otolaryngology, China-Japan Union Hospital of Jilin University, Changchun 130033, P. R. China
  • Received:2012-08-31 Revised:2012-12-16 Online:2013-04-01 Published:2013-03-20
  • Supported by:

    Supported by the National Natural Science Foundation of China(No.30973274) and the Doctoral Fund of Youth Scholars of the Ministry of Education of China(No.20090061120073).

Abstract:

SAV1 is a core component involved in the Hippo pathway which can control the organ size via regulating cell proliferation and apoptosis simultaneously. We explored the regulatory mechanism of SAV1. We established the HEK293T cell pool, the cells in which can stably express SAV1 by retroviruses infection and found that SAV1 stable cells reduced the movement of themselves and resulted in multicellular aggregation. We purified SAV1 interacting protein complex using streptavidin resin and subsequently analyzed the digested peptides by high performance liquid chromatography(HPLC)-MS/MS. Results show that about 150 proteins were identified in the complex of SAV1 with protein. TUBA1A, OTUD4, and ATD were identified as proteins interacting with SAV1. Importantly, PP1A, serine/threonine protein phosphatase PP1-alpha 1 catalytic subunit, was also in the top 10 list. The interaction between PP1A and SAV1 was detected by both co-immunoprecipitation(CO-IP) and immunostaining. Our results indicate that PP1A might be the phosphatase of SAV1 and may take part in the regulation of the Hippo pathway.

Key words: SAV1, PP1A, High performance liquid chromatography(HPLC)-MS/MS, Interacting protein complex