Further Studies on a-Oxidation
Degradation of 2-Hydroxy Fatty Acids & Thiamine Dependence
Acta Biomedica Lovaniensia, No. 409
By Mieke Sniekers
December 2007
Leuven University Press
Distributed By Coronet Books
ISBN: 9789058676528 
115 pages, Illustrated, 6 1/8 x 9 1/2"
$97.50 Paper Original
GENERAL INTRODUCTION
1. Mammalian fatty acid degradation and peroxisomes
2. 3-Methyl-branched and 2- hydroxyl straight chain fatty acids
2.1 Phytanic acid: the major 3-methyl-branched fatty acid
2.1.1 Origin of phytanic acid
2.1.2 Conversion of phytol to phytanic acid
2.1.3 Phytanic acid in the diet
2.2 2-Hydroxy straight chain fatty acids
2.2.1 Occurrence of 2-hydroxy fatty acids
2.2.2 Role of 2-hydroxy fatty acids in brain
3. Alpha-oxidation of 3-methyl-branched fatty acids
3.1 The alpha-oxidation pathway of phytanic acid
3.1.1 Activation and peroxisomal transport of phytanic acid
3.1.2 Hydroxylation by phytanoyl-CoA hydroxylase
3.1.3 Cleavage of the 2-hydroxy intermediate by 2-hydroxyphytanoyl-CoA
lyase
3.1.4 Conversion of pristinal to pristanic acid
3.1.5 Further degradation of pristanic acid by beta-oxidation
3.2 Refsum disease
4. Alpha-oxidation of straight chain fatty acids in the brain
4.1 alpha-hydroxylation of straight chain fatty acids int he brain
4.2 alpha-oxidation of 2-hydroxy fatty acids
5. TPP dependence of alpha-oxidation
AIM AND OUTLINE OF THE STUDY
EXPERIMENTAL PROCEDURES
1. Materials Part 1:alpha-oxidation of 2-hydroxy fatty acids Part 2:Thiamine dependence of mammalian alpha-oxidation REFERENCES Medical Science
1.1 General
1.2 Fatty acids and derivatives
2. Animals, human tissues, cell lines and strains
3. Molecular biology techniques
3.1 Constructs
3.2 Transformation of bacteria and plasmid isolation
3.3 Eukaryotic expression of human acyl-CoA synthetases
3.4 Purification of recombinant human 2-hydroxyphytanoyl-CoA lyase
4. Metabolic studies and analytical procedures
4.1 Preparations of hepatocytes, homogenates, subcellular fractions and cell
lysates
4.2 Oxidation experiments in cells and homogenates
4.3 Enzyme activity measurements
4.3.1 Acyl-CoA synthetase
4.3.2 2-hydroxyphytanoyl-Coa lyase
4.3.3 Pyruvate decarboxylase
RESULTS AND DISCUSSION
1.1 Introduction
1.2 Results and discussion
1.2.1 Acyl-CoA synthetase
1.2.2 Cofactor requirements
1.2.3 Identification of intermediates and end products in homogenates
1.2.4 Search for the acyl-CoA synthetase catalyzing the activation fo 2-hydroxy
fatty acids
1.2.5 Tissue distribution of lyase activity
1.2.6 Subcellular distribution of lyase activity
1.2.7 Identificationof 2-hydroxyphytanoyl-CoA lyase as the cleavage enzyme
in the alpha-oxidation of long chain 2-hydroxy fatty acids
1.2.8 Degradation of very long chain 2-hydroxy fatty acids in intact cells
1.3 Conclusion
2.1 Introduction
2.2 Results and discussion
2.2.1 Effects of thiamine depletion on fatty acid oxidation and TPP-dependent
enzymes
2.2.2 Hepatic fatty acid oxidation and 2-hydroxyacyl-CoA lyase 1 activity
in thiamine-deficient rats
2.2.3 Is alpha-oxidation affected in thiamine-responsive megaloblastic
anemia?
2.3 Conclusion
GENERAL DISCUSSION
SUMMARY / SAMENVATTING
