PMID- 30972724
OWN - NLM
STAT- MEDLINE
DCOM- 20191211
LR  - 20200701
IS  - 1879-1123 (Electronic)
IS  - 1044-0305 (Print)
IS  - 1044-0305 (Linking)
VI  - 30
IP  - 7
DP  - 2019 Jul
TI  - Selenoproteome Identification in Inflamed Murine Primary Bone Marrow-Derived 
      Macrophages by Nano-LC Orbitrap Fusion Tribrid Mass Spectrometry.
PG  - 1276-1283
LID - 10.1007/s13361-019-02192-9 [doi]
AB  - Selenium (Se) functions as a cellular redox gatekeeper through its incorporation 
      into proteins as the 21st amino acid, selenocysteine (Sec). Supplementation of 
      macrophages with exogenous Se (as sodium selenite) downregulates inflammation and 
      intracellular oxidative stress by effectively restoring redox homeostasis upon 
      challenge with bacterial endotoxin lipopolysaccharide (LPS). Here, we examined 
      the use of a standard Tandem Mass Tag (TMT)-labeling mass spectrometry-based 
      proteomic workflow to quantitate and examine temporal regulation of 
      selenoproteins in such inflamed cells. Se-deficient murine primary bone 
      marrow-derived macrophages (BMDMs) exposed to LPS in the presence or absence of 
      selenite treatment for various time periods (0-20 h) were used to analyze the 
      selenoproteome expression using isobaric labeling and shotgun proteomic workflow. 
      To overcome the challenge of identification of Sec peptides, we used the 
      identification of non-Sec containing peptides downstream of Sec as a reliable 
      evidence of ribosome readthrough indicating efficient decoding of Sec codon. 
      Results indicated a temporal regulation of the selenoproteome with a general 
      increase in their expression in inflamed cells in a Se-dependent manner. Selenow, 
      Gpx1, Msrb1, and Selenom were highly upregulated upon stimulation with LPS when 
      compared to other selenoproteins. Interestingly, Selenow appeared to be one 
      amongst the highly regulated selenoproteins in macrophages that was previously 
      thought to be mainly restricted to myocytes. Collectively, TMT-labeling method of 
      non-Sec peptides offers a reliable method to quantitate and study temporal 
      regulation of selenoproteins; however, further optimization to include 
      Sec-peptides could make this strategy more robust and sensitive compared to other 
      semi-quantitative or qualitative methods. Graphical Abstract.
FAU - Korwar, Arvind M
AU  - Korwar AM
AD  - Department of Veterinary and Biomedical Sciences and The Penn State Cancer 
      Institute, The Pennsylvania State University, 115 Henning Building, University 
      Park, PA, 16802, USA.
FAU - Shay, Ashley E
AU  - Shay AE
AD  - Department of Veterinary and Biomedical Sciences and The Penn State Cancer 
      Institute, The Pennsylvania State University, 115 Henning Building, University 
      Park, PA, 16802, USA.
FAU - Basrur, Venkatesha
AU  - Basrur V
AD  - Department of Pathology, Proteomics Research Core Facility, Medical School, 
      University of Michigan, Ann Arbor, MI, USA.
FAU - Conlon, Kevin
AU  - Conlon K
AD  - Department of Pathology, Proteomics Research Core Facility, Medical School, 
      University of Michigan, Ann Arbor, MI, USA.
FAU - Prabhu, K Sandeep
AU  - Prabhu KS
AD  - Department of Veterinary and Biomedical Sciences and The Penn State Cancer 
      Institute, The Pennsylvania State University, 115 Henning Building, University 
      Park, PA, 16802, USA. ksp4@psu.edu.
LA  - eng
GR  - R01 DK077152/DK/NIDDK NIH HHS/United States
GR  - Hatch Fund 4605; Accession # 1010021/USDA-NIFA/
GR  - DK077152/National Institute of Diabetes and Digestive and Kidney Diseases/
PT  - Journal Article
DEP - 20190409
PL  - United States
TA  - J Am Soc Mass Spectrom
JT  - Journal of the American Society for Mass Spectrometry
JID - 9010412
RN  - 0 (Lipopolysaccharides)
RN  - 0 (Selenoproteins)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Inflammation/immunology
MH  - Lipopolysaccharides/immunology
MH  - Macrophages/*chemistry/immunology
MH  - Male
MH  - Mice, Inbred C57BL
MH  - Proteomics/methods
MH  - Selenoproteins/*analysis/immunology
MH  - Tandem Mass Spectrometry/methods
PMC - PMC6592718
MID - NIHMS1526751
OTO - NOTNLM
OT  - Inflammation
OT  - Lipopolysaccharide
OT  - Proteomics
OT  - Redox
OT  - Resolution
EDAT- 2019/04/12 06:00
MHDA- 2019/12/18 06:00
PMCR- 2020/07/01
CRDT- 2019/04/12 06:00
PHST- 2018/11/20 00:00 [received]
PHST- 2019/03/11 00:00 [accepted]
PHST- 2019/02/11 00:00 [revised]
PHST- 2019/04/12 06:00 [pubmed]
PHST- 2019/12/18 06:00 [medline]
PHST- 2019/04/12 06:00 [entrez]
PHST- 2020/07/01 00:00 [pmc-release]
AID - 10.1007/s13361-019-02192-9 [pii]
AID - 10.1007/s13361-019-02192-9 [doi]
PST - ppublish
SO  - J Am Soc Mass Spectrom. 2019 Jul;30(7):1276-1283. doi: 
      10.1007/s13361-019-02192-9. Epub 2019 Apr 9.