Hanson B J, Carrozzo R, Piemonte F, Tessa A, Robinson B H, Capaldi R A
Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403, USA.
J Biol Chem. 2001 May 11;276(19):16296-301. doi: 10.1074/jbc.M011162200. Epub 2001 Feb 7.
Cytochrome c oxidase (COX) deficiency is the most common respiratory chain defect in childhood and is clinically heterogeneous. We report a study of six patients with COX deficiencies. Two of the patients had as yet undefined defects, three patients had Surf-1 mutations, and one patient had a 15-base pair deletion in the COX III subunit. We show that quantitative measurements of steady-state levels of subunits by monoclonal antibody reactivity, when used in combination with a discontinuous sucrose gradient methods, provide an improved diagnosis of COX deficiencies by distinguishing between kinetic, stability, and assembly defects. The two mutants of undefined etiology had a full complement of subunits with one stable and the other partially unstable to detergent solubilization. Both are likely to carry mutations in nuclear-encoded subunits of the complex. The three Surf-1 mutants and the COX III mutant each had reduced steady-state levels of subunits but variable associations of the residual subunits. This information, as well as aiding in diagnosis, helps in understanding the genotype-phenotype relationships of COX deficiencies and provides insight into the mechanism of assembly of the enzyme complex.
细胞色素c氧化酶(COX)缺乏症是儿童期最常见的呼吸链缺陷,临床症状具有异质性。我们报告了一项对6例COX缺乏症患者的研究。其中2例患者存在尚未明确的缺陷,3例患者存在Surf-1突变,1例患者的COX III亚基有15个碱基对的缺失。我们发现,当结合不连续蔗糖梯度法,通过单克隆抗体反应性对亚基稳态水平进行定量测量时,能够通过区分动力学、稳定性和组装缺陷来改进COX缺乏症的诊断。病因不明的两个突变体具有完整的亚基互补,其中一个对去污剂溶解稳定,另一个部分不稳定。两者可能都携带该复合物核编码亚基的突变。三个Surf-1突变体和COX III突变体的亚基稳态水平均降低,但残余亚基的结合情况各不相同。这些信息不仅有助于诊断,还有助于理解COX缺乏症的基因型-表型关系,并为酶复合物的组装机制提供见解。