The Importance of Calcium Ions for Determining Mitochondrial Glycerol-3-Phosphate Dehydrogenase Activity When Measuring Uncoupling Protein 1 (UCP1) Function in Mitochondria Isolated from Brown Adipose Tissue

2018 ◽  
pp. 325-336 ◽  
Author(s):  
Kieran J. Clarke ◽  
Richard K. Porter
Keyword(s):  
Adipose Tissue ◽  
Brown Adipose Tissue ◽  
Dehydrogenase Activity ◽  
Calcium Ions ◽  
Uncoupling Protein ◽  
Uncoupling Protein 1 ◽  
Brown Adipose ◽  
Glycerol 3 Phosphate Dehydrogenase

scholarly journals Uncoupling protein 1 binds one nucleotide per monomer and is stabilized by tightly bound cardiolipin

2015 ◽  
Vol 112 (22) ◽  
pp. 6973-6978 ◽  
Author(s):  
Yang Lee ◽  
Chrissie Willers ◽  
Edmund R. S. Kunji ◽  
Paul G. Crichton
Keyword(s):  
Adipose Tissue ◽  
Brown Adipose Tissue ◽  
Uncoupling Protein ◽  
Size Exclusion ◽  
Uncoupling Protein 1 ◽  
Mitochondrial Carriers ◽  
Mitochondrial Inner Membrane ◽  
Brown Adipose ◽  
Integral Role ◽  
Covalent Chromatography

Uncoupling protein 1 (UCP1) catalyzes fatty acid-activated, purine nucleotide-sensitive proton leak across the mitochondrial inner membrane of brown adipose tissue to produce heat, and could help combat obesity and metabolic disease in humans. Studies over the last 30 years conclude that the protein is a dimer, binding one nucleotide molecule per two proteins, and unlike the related mitochondrial ADP/ATP carrier, does not bind cardiolipin. Here, we have developed novel methods to purify milligram amounts of UCP1 from native sources by using covalent chromatography that, unlike past methods, allows the protein to be prepared in defined conditions, free of excess detergent and lipid. Assessment of purified preparations by TLC reveal that UCP1 retains tightly bound cardiolipin, with a lipid phosphorus content equating to three molecules per protein, like the ADP/ATP carrier. Cardiolipin stabilizes UCP1, as demonstrated by reconstitution experiments and thermostability assays, indicating that the lipid has an integral role in the functioning of the protein, similar to other mitochondrial carriers. Furthermore, we find that UCP1 is not dimeric but monomeric, as indicated by size exclusion analysis, and has a ligand titration profile in isothermal calorimetric measurements that clearly shows that one nucleotide binds per monomer. These findings reveal the fundamental composition of UCP1, which is essential for understanding the mechanism of the protein. Our assessment of the properties of UCP1 indicate that it is not unique among mitochondrial carriers and so is likely to use a common exchange mechanism in its primary function in brown adipose tissue mitochondria.

Tea catechins enhance the mRNA expression of uncoupling protein 1 in rat brown adipose tissue

2008 ◽  
Vol 19 (12) ◽  
pp. 840-847 ◽  
Author(s):  
Sachiko Nomura ◽  
Takashi Ichinose ◽  
Manabu Jinde ◽  
Yu Kawashima ◽  
Kaoru Tachiyashiki ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Brown Adipose Tissue ◽  
Mrna Expression ◽  
Uncoupling Protein ◽  
Uncoupling Protein 1 ◽  
Tea Catechins ◽  
Brown Adipose

scholarly journals Differential regulation of uncoupling protein-1, -2 and -3 gene expression by sympathetic innervation in brown adipose tissue of thermoneutral or cold-exposed rats

FEBS Letters ◽  
1999 ◽  
Vol 444 (2-3) ◽  
pp. 181-185 ◽  
Author(s):  
Frédérique Denjean ◽  
Joël Lachuer ◽  
Alain Géloën ◽  
Frédérique Cohen-Adad ◽  
Colette Moulin ◽  
...  
Keyword(s):  
Gene Expression ◽  
Adipose Tissue ◽  
Brown Adipose Tissue ◽  
Uncoupling Protein ◽  
Sympathetic Innervation ◽  
Differential Regulation ◽  
Uncoupling Protein 1 ◽  
Brown Adipose

scholarly journals Uncoupling protein-1 (UCP1) contributes to the basal proton conductance of brown adipose tissue mitochondria

2009 ◽  
Vol 41 (4) ◽  
pp. 335-342 ◽  
Author(s):  
Nadeene Parker ◽  
Paul G. Crichton ◽  
Antonio J. Vidal-Puig ◽  
Martin D. Brand
Keyword(s):  
Adipose Tissue ◽  
Brown Adipose Tissue ◽  
Uncoupling Protein ◽  
Proton Conductance ◽  
Uncoupling Protein 1 ◽  
Brown Adipose
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