Elucidating the Catalytic Subunit Composition of Distinct Proteasome Subtypes: A Crosslinking Approach Employing Bifunctional Activity-Based Probes

作者：Carmony Kimberly Cornish; Sharma Lalit Kumar; Lee Do Min; Park Ji Eun; Lee Wooin^*; Kim Kyung Bo

来源：ChemBioChem, 2015, 16(2): 284-292.

DOI：10.1002/cbic.201402491

• constitutive proteasome
• crosslinking agents
• immunoproteasome
• intermediate proteasome
• proteasome subtype

In addition to two well-recognized proteasome subtypes-constitutive proteasomes and immunoproteasomes-mounting evidence also suggests the existence of intermediate proteasome subtypes containing unconventional mixtures of catalytic subunits. Although they appear to play unique biological roles, the lack of practical methods for detecting distinct proteasome subtypes has limited functional investigations. Here, we report the development of activity-based probes that cross-link two catalytic subunits within intact proteasome complexes. Identification of the crosslinked subunit pairs provides direct evidence of the catalytic subunit composition of proteasomes. Using these probes, we found that U266 multiple myeloma cells contain intermediate proteasomes comprising both beta 1i and beta 2, but not beta 1 and beta 2i, consistent with previous findings with other cell types. Our bifunctional probes can be utilized in functional investigations of distinct proteasome subtypes in various biological settings.

• 出版日期2015-1-19