Ameis D, Kobayashi J, Davis R C, Ben-Zeev O, Malloy M J, Kane J P, Lee G, Wong H, Havel R J, Schotz M C
Veterans Administration Wadsworth Medical Center, Los Angeles, California 90073.
J Clin Invest. 1991 Apr;87(4):1165-70. doi: 10.1172/JCI115114.
Complete deficiency of lipoprotein lipase (LPL) causes the chylomicronemia syndrome. To understand the molecular basis of LPL deficiency, two siblings with drastically reduced postheparin plasma lipolytic activities were selected for analysis of their LPL gene. We used the polymerase chain reaction to examine the nine coding LPL exons in the two affected siblings and three relatives. DNA sequence analysis revealed a single nucleotide change compared with the normal LPL cDNA: a G----A substitution at nucleotide position 680. This transition caused a replacement of glutamic acid for glycine at amino acid residue 142 of the mature LPL protein. Amino acid sequence comparisons of the region surrounding glycine-142 indicated that it is highly conserved among lipases from different species, suggesting a crucial role of this domain for the LPL structure. Expression studies of the mutant LPL cDNA in COS-7 cells produced normal amounts of enzyme mass. However, the mutated LPL was not catalytically active, nor was it efficiently secreted from the cells. This established that the Gly----Glu substitution at amino acid 142 is sufficient to abolish enzymatic activity and to result in the chylomicronemia syndrome observed in these patients.
脂蛋白脂肪酶(LPL)完全缺乏会导致乳糜微粒血症综合征。为了解LPL缺乏的分子基础,选择了两名肝素后血浆脂解活性大幅降低的同胞进行LPL基因分析。我们使用聚合酶链反应检测了两名患病同胞和三名亲属的LPL九个编码外显子。DNA序列分析显示,与正常LPL cDNA相比有一个单核苷酸变化:核苷酸位置680处的G→A替换。这种转换导致成熟LPL蛋白的第142位氨基酸残基处的甘氨酸被谷氨酸取代。对甘氨酸-142周围区域的氨基酸序列比较表明,它在来自不同物种的脂肪酶中高度保守,表明该结构域对LPL结构起着关键作用。在COS-7细胞中对突变型LPL cDNA进行的表达研究产生了正常量的酶量。然而,突变的LPL没有催化活性,也没有从细胞中有效分泌。这证实了第142位氨基酸处的甘氨酸→谷氨酸替换足以消除酶活性,并导致在这些患者中观察到的乳糜微粒血症综合征。