open access

Vol 77, No 2 (2009)
ORIGINAL PAPERS
Published online: 2009-03-16
Submitted: 2013-02-22
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Application of Western blotting for the detection of uncoupling protein-2 (UCP-2) in mitochondria from smokers and non-smokers

Kinga I. Stańczak-Mrozek, Monika Kolanowska, Paweł Grieb
Pneumonol Alergol Pol 2009;77(2):118-122.

open access

Vol 77, No 2 (2009)
ORIGINAL PAPERS
Published online: 2009-03-16
Submitted: 2013-02-22

Abstract


Introduction: 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.
Material and 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.
Conclusion: 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.

Abstract


Introduction: 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.
Material and 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.
Conclusion: 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.
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Keywords

uncoupling protein-2; lymphocytes; mitochondria; tobacco smoking; Western blotting

About this article
Title

Application of Western blotting for the detection of uncoupling protein-2 (UCP-2) in mitochondria from smokers and non-smokers

Journal

Advances in Respiratory Medicine

Issue

Vol 77, No 2 (2009)

Pages

118-122

Published online

2009-03-16

Bibliographic record

Pneumonol Alergol Pol 2009;77(2):118-122.

Keywords

uncoupling protein-2
lymphocytes
mitochondria
tobacco smoking
Western blotting

Authors

Kinga I. Stańczak-Mrozek
Monika Kolanowska
Paweł Grieb

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