McGovern M M, Mandell N, Haskins M, Desnick R J
Genetics. 1985 Aug;110(4):733-49. doi: 10.1093/genetics/110.4.733.
The identification of a second structural gene mutation at the feline arylsulfatase B locus (MPS VIb) provided the opportunity to investigate the expression of allelism by characterization of the residual enzymatic activity in feline mucopolysaccharidosis VI, an animal analogue of human Maroteaux-Lamy syndrome. Matings were designed to produce affected homozygotes who were homoallelic for the MPS VIa and MPS VIb mutations or heteroallelic genetic compounds (MPS VIa/VIb). The physicokinetic and immunological properties of the partially purified residual hepatic arylsulfatase B isozymes from the affected homoallelic and heteroallelic cats were compared to those of the normal feline enzyme. The residual hepatic arylsulfatase B activities from the inbred MPS VIa and MPS VIb homozygotes were distinguished by differences in physicokinetic and immunological properties. The newly identified mutant isozyme b had abnormal kinetic properties toward artificial and natural substrates, normal cryo- and thermostabilities, a normal molecular weight and an altered electrophoretic mobility. Polyacrylamide gel electrophoresis demonstrated that the mutant b subunits formed dimers with normal subunits in obligate heterozygotes (MPS VI+/b). In contrast, mutant isozyme a subunits from obligate MPS VIa/+ heterozygotes did not dimerize with the normal subunit, and the mutant and normal isozymes could be separated by anion exchange chromatography and polyacrylamide gel electrophoresis. Characterization of the partially purified residual hepatic arylsulfatase B activity from the heteroallelic homozygotes revealed the presence of both mutant isozymes a and b. The demonstration of two allelic mutations in the feline arylsulfatase B gene documented the occurrence of genetic heterogeneity in feline mucopolysaccharidosis VI and permitted characterization of the enzymatic defect in homoallelic and heteroallelic (genetic compound) homozygotes, providing a model for the study of allelism in human genetic disorders.
在猫芳基硫酸酯酶B位点(MPS VIb)发现的第二个结构基因突变,为通过对猫黏多糖贮积症VI(人类马罗-拉米综合征的动物类似物)中残留酶活性的表征来研究等位基因的表达提供了机会。设计交配方案以产生受影响的纯合子,这些纯合子对于MPS VIa和MPS VIb突变是同型等位基因,或者是异型等位基因遗传化合物(MPS VIa/VIb)。将来自受影响的同型等位基因和异型等位基因猫的部分纯化的残留肝脏芳基硫酸酯酶B同工酶的物理动力学和免疫学特性与正常猫酶的特性进行了比较。来自近交MPS VIa和MPS VIb纯合子的残留肝脏芳基硫酸酯酶B活性通过物理动力学和免疫学特性的差异得以区分。新鉴定的突变同工酶b对人工和天然底物具有异常的动力学特性,具有正常的冷冻和热稳定性,正常的分子量以及改变的电泳迁移率。聚丙烯酰胺凝胶电泳表明,突变的b亚基在纯合杂合子(MPS VI+/b)中与正常亚基形成二聚体。相反,来自纯合MPS VIa/+杂合子的突变同工酶a亚基不与正常亚基二聚化,并且突变和正常同工酶可以通过阴离子交换色谱法和聚丙烯酰胺凝胶电泳分离。对来自异型等位基因纯合子的部分纯化的残留肝脏芳基硫酸酯酶B活性的表征揭示了突变同工酶a和b的存在。猫芳基硫酸酯酶B基因中两个等位基因突变的证明记录了猫黏多糖贮积症VI中遗传异质性的发生,并允许对同型等位基因和异型等位基因(遗传化合物)纯合子中的酶缺陷进行表征,为研究人类遗传疾病中的等位基因提供了一个模型。