[Application of Western blotting for the detection of uncoupling protein-2 (UCP-2) in mitochondria from smokers and non-smokers].
BACKGROUND
Uncoupling proteins (UCPs) are a family of transmembrane anion transporters present in the inner mitochondrial membrane. UCP-2, which exhibits the widest distribution in various tissues, plays an important role in many physiological processes. Human UCP-2 studies have been hampered by the lack of a method for measuring this protein in an easily accessible human tissue, e.g. blood. The aim of this study was to develop such a method and test its utility by comparing UCP-2 levels in smokers and non-smokers.


METHODS
Venous blood samples from 10 smoking and seven non-smoking volunteers were used for the study; lymphocytes were isolated employing Lymphoprep. UCP-2 levels were measured by Western blotting combined with chemoluminescence detection.


RESULTS
Total lymphocyte homogenates were found useless for measuring UCP-2 levels, but it was possible to measure UCP-2 in homogenates of purified lymphocyte mitochondria. There was a significant, though moderate, linear correlation between UCP-2 level and daily cigarette use. UCP-2 level in peripheral blood lymphocytes from smokers was higher than that in non-smokers.


CONCLUSIONS
The method for measuring UCP-2 in peripheral blood lymphocytes opens the possibility of UCP-2 screening studies in humans and thus may be useful for studying the role of the protein in human physiology and pathology.
Stańczak-Mrozek
Kinga I
KI
Zakład Farmakologii Doświadczalnej Instytutu Medycyny Doświadczalnej i Klinicznej PAN w Warszawie. kingastanczak@wp.pl
Kolanowska
Monika
M
Grieb
Paweł
P
D000328:Adult; D015153:Blotting, Western / Q000379:methods; D005260:Female; D006801:Humans; D007473:Ion Channels / Q000097:blood*; D008297:Male; D008875:Middle Aged; D008928:Mitochondria / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000097:blood*; D015999:Multivariate Analysis; D012044:Regression Analysis; D012907:Smoking / Q000097:blood*; D014029:Tobacco Use Disorder / Q000097:blood*; D014475:Uncoupling Agents / Q000097:blood*; D000071246:Uncoupling Protein 2
D007473:Ion Channels; D024101:Mitochondrial Proteins; C000604389:UCP2 protein, human; D014475:Uncoupling Agents; D000071246:Uncoupling Protein 2