Open AccessRevealing the cellular degradome by mass spectrometry analysis of proteasome-cleaved peptides
Author(s) -
Hila Wolf-Levy,
Aaron Javitt,
Avital EisenbergLerner,
Assaf Kacen,
Adi Ulman,
Daoud Sheban,
Bareket Dassa,
Vered Fishbain-Yoskovitz,
Carmelo Carmona-Rivera,
Matthias P. Kramer,
Neta Nudel,
Ifat Regev,
Liron Zahavi,
Dalia Elinger,
Mariana J. Kaplan,
David Morgenstern,
Yishai Levin,
Yifat Merbl
Publication year - 2018
Publication title -
nature biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 15.358
H-Index - 445
eISSN - 1546-1696
pISSN - 1087-0156
DOI - 10.1038/nbt.4279
Subject(s) - proteasome , immune system , proteolysis , chemistry , protein degradation , microbiology and biotechnology , biochemistry , biology , immunology , enzyme
Cellular function is critically regulated through degradation of substrates by the proteasome. To enable direct analysis of naturally cleaved proteasomal peptides under physiological conditions, we developed mass spectrometry analysis of proteolytic peptides (MAPP), a method for proteasomal footprinting that allows for capture, isolation and analysis of proteasome-cleaved peptides. Application of MAPP to cancer cell lines as well as primary immune cells revealed dynamic modulation of the cellular degradome in response to various stimuli, such as proinflammatory signals. Further, we performed analysis of minute amounts of clinical samples by studying cells from the peripheral blood of patients with systemic lupus erythematosus (SLE). We found increased degradation of histones in patient immune cells, thereby suggesting a role of aberrant proteasomal degradation in the pathophysiology of SLE. Thus, MAPP offers a broadly applicable method to facilitate the study of the cellular-degradation landscape in various cellular conditions and diseases involving changes in proteasomal degradation, including protein aggregation diseases, autoimmunity and cancer.