New insights into the pathological mechanisms of cerebrotendinous xanthomatosis in the Taiwanese using genomic and proteomic tools.
Cerebrotendinous xanthomatosis (CTX) is an autosomal recessive lipid-storage disorder caused by a deficiency of the mitochondrial sterol 27-hydroxylase. Genetic analysis utilizing SSCP and direct DNA sequencing identified a new mutation. One base-pair of cytosine was deleted at codon 326 on exon 2 of CYP27 in all CTX patients while their father was heterozygotic. This novel point deletion predicts a frameshift in mRNA (Pro(102) -->Leu) and results in the appearance of a premature termination codon (TGA) to substitute for Val(106) (GTG). To characterize the pathological mechanism of CTX patients, the protein profiles of serum and leukocytes extracted from these subjects were presented by means of proteomic technologies including 2-DE and MALDI-TOF analysis. According to the results, the amount of vinculin, ABP-280, talin and vimentin in leukocytes of CTX patients had changed significantly, reflecting the changes in membrane dynamics concerning cholestanol accumulation. The expression of target proteins in CTX patients and control was further verified by western blotting which indicated the same tendency as 2-DE data. This is the first paper to integrate both genomic and proteomic concepts for analyzing the possible mechanism of CTX and provides more information for related study in the future.
Wang
Pei-Weg
PW
Department of Biological Sciences, National Sun Yat-Sen University, Kaohsiung, Taiwan.
Chang
Wen-Neng
WN
Lu
Chen-Hsin
CH
Chao
David
D
Schrag
Christiaan
C
Pan
Tai-Long
TL
D000595:Amino Acid Sequence; D001483:Base Sequence; D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D002648:Child; D053493:Cholestanetriol 26-Monooxygenase; D004247:DNA / Q000097:blood / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D023281:Genomics; D006801:Humans; D007962:Leukocytes / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D010375:Pedigree; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D040901:Proteomics; D012689:Sequence Homology, Nucleic Acid; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013250:Steroid Hydroxylases / Q000235:genetics / Q000378:metabolism; D013624:Taiwan; D019294:Xanthomatosis, Cerebrotendinous / Q000235:genetics / Q000378:metabolism / Q000473:pathology
D015415:Biomarkers; D004247:DNA; D013250:Steroid Hydroxylases; C506831:CYP27A1 protein, human; D053493:Cholestanetriol 26-Monooxygenase