Detecting Posttranslational Modifications of Hsp90

Rebecca A Sager; Mark R Woodford; Len Neckers; Mehdi Mollapour

doi:10.1007/978-1-4939-7477-1_16

Detecting Posttranslational Modifications of Hsp90

Methods Mol Biol. 2018:1709:209-219. doi: 10.1007/978-1-4939-7477-1_16.

Authors

Rebecca A Sager^{1

2

3}, Mark R Woodford^{1

2}, Len Neckers⁴, Mehdi Mollapour^{5

6

7}

Affiliations

¹ Department of Urology, SUNY Upstate Medical University, 750 E. Adams Street, Syracuse, NY, 13210, USA.
² Upstate Cancer Center, SUNY Upstate Medical University, Syracuse, NY, USA.
³ Department of Biochemistry and Molecular Biology, SUNY Upstate Medical University, 750 E. Adams Street, Syracuse, NY, 13210, USA.
⁴ Urologic Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, 9000 Rockville Pike, Bethesda, MD, 20892, USA.
⁵ Department of Urology, SUNY Upstate Medical University, 750 E. Adams Street, Syracuse, NY, 13210, USA. mollapom@upstate.edu.
⁶ Upstate Cancer Center, SUNY Upstate Medical University, Syracuse, NY, USA. mollapom@upstate.edu.
⁷ Department of Biochemistry and Molecular Biology, SUNY Upstate Medical University, 750 E. Adams Street, Syracuse, NY, 13210, USA. mollapom@upstate.edu.

Abstract

The molecular chaperone Heat Shock Protein 90 (Hsp90) is essential in eukaryotes. Hsp90 chaperones proteins that are important determinants of multistep carcinogenesis. The chaperone function of Hsp90 is linked to its ability to bind and hydrolyze ATP. Co-chaperones as well as posttranslational modifications (phosphorylation, SUMOylation, and ubiquitination) are important for its stability and regulation of the ATPase activity. Both mammalian and yeast cells can be used to express and purify Hsp90 and also detect its posttranslational modifications by immunoblotting.

Keywords: Heat shock protein 90 (Hsp90); Molecular chaperones; Phosphorylation; Posttranslational modification; SUMOylation; Ubiquitination.

MeSH terms

Acetylation
Animals
Blotting, Western / methods*
HSP90 Heat-Shock Proteins / analysis*
HSP90 Heat-Shock Proteins / chemistry
HSP90 Heat-Shock Proteins / metabolism*
Humans
Phosphorylation
Protein Processing, Post-Translational*
Saccharomyces cerevisiae / metabolism
Ubiquitination

Substances

HSP90 Heat-Shock Proteins

Grants and funding

R01 GM124256/GM/NIGMS NIH HHS/United States