Research output: Contribution to journal › Article › peer-review
Proteomic analysis of affinity-purified extracellular proteasomes reveals exclusively 20S complexes. / Kulichkova, Valentina A.; Artamonova, Tatiana O.; Lyublinskaya, Olga G.; Khodorkovskii, Mikhail A.; Tomilin, Alexey N.; Tsimokha, Anna S.
In: Oncotarget, Vol. 8, No. 60, 24.11.2017, p. 102134-102149.Research output: Contribution to journal › Article › peer-review
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TY - JOUR
T1 - Proteomic analysis of affinity-purified extracellular proteasomes reveals exclusively 20S complexes
AU - Kulichkova, Valentina A.
AU - Artamonova, Tatiana O.
AU - Lyublinskaya, Olga G.
AU - Khodorkovskii, Mikhail A.
AU - Tomilin, Alexey N.
AU - Tsimokha, Anna S.
PY - 2017/11/24
Y1 - 2017/11/24
N2 - Proteasome-mediated proteolysis is important for many basic cellular processes. In addition to their functions in the cell, proteasomes have been found in physiological fluids of both healthy and diseased humans including cancer patients. Higher levels of these proteasomes are associated with higher cancer burden and stage. The etiology and functions of these proteasomes, referred to as circulating, plasmatic, or extracellular proteasomes (ex-PSs), are unclear. Here we show that human cancer cell lines, as well as human endometrium-derived mesenchymal stem cells (hMESCs), release proteasome complexes into culture medium (CM). To define ex-PS composition, we have affinity purified them from CM conditioned by human leukemia cell line K562. Using matrix-assisted laser desorption/ionization (MALDI) Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry (MS), we have identified core 20S proteasome subunits and a set of 15 proteasome-interacting proteins (PIPs), all previously described as exosome cargo proteins. Three of them, PPIase A, aldolase A, and transferrin, have never been reported as PIPs. The study provides compelling arguments that ex-PSs do not contain 19S or PA200 regulatory particles and are represented exclusively by the 20S complex.
AB - Proteasome-mediated proteolysis is important for many basic cellular processes. In addition to their functions in the cell, proteasomes have been found in physiological fluids of both healthy and diseased humans including cancer patients. Higher levels of these proteasomes are associated with higher cancer burden and stage. The etiology and functions of these proteasomes, referred to as circulating, plasmatic, or extracellular proteasomes (ex-PSs), are unclear. Here we show that human cancer cell lines, as well as human endometrium-derived mesenchymal stem cells (hMESCs), release proteasome complexes into culture medium (CM). To define ex-PS composition, we have affinity purified them from CM conditioned by human leukemia cell line K562. Using matrix-assisted laser desorption/ionization (MALDI) Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry (MS), we have identified core 20S proteasome subunits and a set of 15 proteasome-interacting proteins (PIPs), all previously described as exosome cargo proteins. Three of them, PPIase A, aldolase A, and transferrin, have never been reported as PIPs. The study provides compelling arguments that ex-PSs do not contain 19S or PA200 regulatory particles and are represented exclusively by the 20S complex.
KW - extracellular proteasome
KW - human leukemia K562 cells
KW - proteasome interacting protein ( PIP)
KW - affinity purification
KW - mass spectrometry
KW - MASS-SPECTROMETRIC ANALYSIS
KW - CIRCULATING PROTEASOMES
KW - 26S PROTEASOME
KW - CANCER CELLS
KW - DNA-REPAIR
KW - UBIQUITIN
KW - PROTEINS
KW - SPERMATOGENESIS
KW - DAMAGE
KW - YEAST
U2 - 10.18632/oncotarget.22230
DO - 10.18632/oncotarget.22230
M3 - статья
VL - 8
SP - 102134
EP - 102149
JO - Oncotarget
JF - Oncotarget
SN - 1949-2553
IS - 60
ER -
ID: 50701056