Ohura T, Miyabayashi S, Narisawa K, Tada K
Department of Pediatrics, Tohoku University School of Medicine, Sendai, Japan.
Hum Genet. 1991 May;87(1):41-4. doi: 10.1007/BF01213089.
Propionic acidemia is an autosomal recessive metabolic disease resulting from a deficiency of propionyl CoA carboxylase (PCC) activity. We have analyzed the molecular heterogeneity of Japanese propionic acidemia patients using anti-human PCC antiserum and cDNA clones coding for the two protein subunits (alpha and beta) of the enzyme. The steady state levels of both alpha and beta subunits of PCC from 15 Japanese patients were determined by Western blot. Three patients had neither alpha nor beta subunits, and the amounts of both alpha and beta subunits were low in 3 other patients. According to our previous data, we classified these 6 patients as having alpha subunit deficiency. In the remaining 8 patients, alpha subunits were normal, but the beta subunits were aberrant. Two patients had low levels of normal-sized beta subunits and 6 had beta subunits smaller than normal in size and greatly reduced in quantity. These 8 patients were assigned to the beta subunit deficiency category. One patient had apparently normal alpha and beta subunits. We could not determine this patient's primary defect. These data reveal the genetic heterogeneity of molecular defects causing propionic acidemia in the Japanese. Southern blot analysis did not reveal any gross alteration in gene structure when DNA was digested with HindIII, EcoRI and TaqI. However, DNA from 3 beta-subunit-deficient patients, when digested with MspI and probed with beta PCC cDNA, revealed a unique 2.7-kb band not observed in blots of DNA from any other patient or 15 normal controls. We conclude that this altered MspI restriction map is the result of a mutation in the beta subunit gene of these patients.
丙酸血症是一种常染色体隐性代谢疾病,由丙酰辅酶A羧化酶(PCC)活性缺乏引起。我们使用抗人PCC抗血清和编码该酶两个蛋白质亚基(α和β)的cDNA克隆,分析了日本丙酸血症患者的分子异质性。通过蛋白质印迹法测定了15名日本患者的PCCα和β亚基的稳态水平。3名患者既没有α亚基也没有β亚基,另外3名患者的α和β亚基含量均较低。根据我们之前的数据,我们将这6名患者归类为α亚基缺乏症。在其余8名患者中,α亚基正常,但β亚基异常。2名患者的正常大小β亚基水平较低,6名患者的β亚基比正常尺寸小且数量大幅减少。这8名患者被归类为β亚基缺乏症。1名患者的α和β亚基明显正常。我们无法确定该患者的主要缺陷。这些数据揭示了导致日本丙酸血症的分子缺陷的遗传异质性。当用HindIII、EcoRI和TaqI消化DNA时,Southern印迹分析未发现基因结构有任何明显改变。然而,来自3名β亚基缺乏症患者的DNA在用MspI消化并用βPCC cDNA探针检测时,显示出一条独特的2.7kb条带,在其他患者或15名正常对照的DNA印迹中未观察到。我们得出结论,这种改变的MspI限制性图谱是这些患者β亚基基因突变的结果。