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标题：New lysosomal acid lipase gene mutants explain the phenotype of Wolman disease and cholesteryl ester storage disease
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作者：Franco Pagani ; Rajalakshmi Pariyarath ; Rodolfo Garcia 等
期刊名称：JLR Papers In Press
印刷版ISSN：0022-2275
电子版ISSN：1539-7262
出版年度：1998
卷号：39
期号：7
页码：1382-1388
语种：English
出版社：American Society for Biochemistry and Molecular Biology
摘要：Deficiency of lysosomal acid lipase (LAL) leads to either Wolman disease (WD) or the more benign cholesteryl ester storage disease (CESD). To identify the molecular basis of the different phenotypes we have characterised the LAL gene mutations in three new patients with LAL deficiency. A patient with WD was homozygote for a null allele Y303X. The other two patients, with CESD, presented either homozygosity for T267I or compound heterozygosity consisting of Q64R and an exon 8 donor splice site substitution (G→A in position –1). The mutants T267I and Q64R and the previously reported L273S, G66V, and H274Y CESD substitutions, overexpressed in stable clones, were found to be fully glycosylated and show an enzymatic activity of 3–8% of that of normal LAL. On the other hand, the Δ254–277 mutant protein derived from exon 8 skipping and the Y303X protein were totally inactive. By transient transfection of hybrid minigene constructs, the CESD G→A (–1) substitution resulted in partial exon inclusion, thus allowing the production of a small amount of normal LAL mRNA and hence of a functional enzyme. In contrast, a G→A substitution observed in WD at position +1 of the same exon 8 donor site resulted in complete exon skipping and the sole production of an inactive Δ254–277 protein. In conclusion, LAL genotypes determine the level of residual enzymatic activity, thus explaining the severity of the phenotype.—Pagani, F., R. Pariyarath, R. Garcia, C. Stuani, A. B. Burlina, G. Ruotolo, M. Rabusin, and F. E. Baralle. New lysosomal acid lipase gene mutants explain the phenotype of Wolman disease and cholesteryl ester storage disease. J. Lipid Res. 1998. 39: 1382–1388.
关键词：alternative splicing ; mutation ; Wolman disease/etiology ; recombinant proteins ; phenotype