Subunit specific monoclonal antibodies show different steady-state levels of various cytochrome-c oxidase subunits in chronic progressive external ophthalmoplegia

被引:53
作者
Taanman, JW
Burton, MD
Marusich, MF
Kennaway, NG
Capaldi, RA
机构
[1] UNIV OREGON, INST MOLEC BIOL, EUGENE, OR 97403 USA
[2] OREGON HLTH SCI UNIV, DEPT MOLEC & MED GENET, PORTLAND, OR 97201 USA
[3] UNIV OREGON, INST NEUROSCI, EUGENE, OR 97403 USA
来源
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR BASIS OF DISEASE | 1996年 / 1315卷 / 03期
关键词
cytochrome c oxidase; mitochondrial protein; immunohistochemistry; monoclonal antibody; chronic progressive external ophthahmoplegia; (human);
D O I
10.1016/0925-4439(95)00127-1
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Monoclonal antibodies recognizing the mitochondrially encoded subunits I and II, and the nuclear-encoded subunits IV, Va, Vb and VIc of human cytochrome-e oxidase were generated. These antibodies are highly specific and allow the assessment of subunit steady-state levels in crude cell extracts and tissue sections. In the experimental human cell line 143B206, which is devoid of mitochondrial DNA, immunovisualization with the antibodies revealed that the nuclear-encoded subunits IV and Va were present in amounts close to that of the parental cell line despite the absence of the mitochondrially encoded subunits. In contrast, the nuclear-encoded subunits Vb and VIc were severely reduced in cell line 143B206, suggesting that unassembled nuclear-encoded subunits are degraded at different rates. In skeletal muscle sections of a patient with chronic progressive external ophthalmoplegia known to harbor the 'common deletion' in a subpopulation of her mitochondrial DNA, most cytochrome-e oxidase activity negative fibers had greatly reduced levels of subunits I, II, Va, Vb and Vie of cytochrome-c oxidase. The steady-state level of subunit IV, however, was less affected. This was particularly evident in cytochrome-c oxidase activity negative fibers with accumulated mitochondria ('ragged-red' fibers) where immunodetection with anti-subunit IV resulted in intense staining. The data presented in this paper demonstrate that the battery of monoclonal antibodies can be employed for diagnostic purposes to analyze steady-state levels of mitochondrially and nuclear-encoded subunits of cytochrome-c oxidase.
引用
收藏
页码:199 / 207
页数:9
相关论文
共 63 条
[11]  
CUNNING WJK, 1994, COLOR ATLAS MUSCLE P, P15
[12]   CYTOCHROME-C-OXIDASE DEFICIENCY [J].
DIMAURO, S ;
LOMBES, A ;
NAKASE, H ;
MITA, S ;
FABRIZI, GM ;
TRITSCHLER, HJ ;
BONILLA, E ;
MIRANDA, AF ;
DEVIVO, DC ;
SCHON, EA .
PEDIATRIC RESEARCH, 1990, 28 (05) :536-541
[13]  
DUBOWITZ V, 1985, MUSCLE BIOPSY PRACTI, P56
[14]   RAPID EXAMINATION OF MUSCLE TISSUE - AN IMPROVED TRICHROME METHOD FOR FRESH-FROZEN BIOPSY SECTIONS [J].
ENGEL, WK ;
CUNNINGHAM, GG .
NEUROLOGY, 1963, 13 (11) :919-&
[15]  
EVANS MJ, 1989, J BIOL CHEM, V264, P14361
[16]   NRF-1 - A TRANSACTIVATOR OF NUCLEAR-ENCODED RESPIRATORY GENES IN ANIMAL-CELLS [J].
EVANS, MJ ;
SCARPULLA, RC .
GENES & DEVELOPMENT, 1990, 4 (06) :1023-1034
[17]  
HAO HL, 1995, AM J HUM GENET, V56, P1017
[18]  
HATEFI Y, 1985, ANNU REV BIOCHEM, V54, P1015, DOI 10.1146/annurev.bi.54.070185.005055
[19]   MOLECULAR-CLONING AND CHARACTERIZATION OF THE RAT CYTOCHROME-C-OXIDASE SUBUNIT VB GENE [J].
HOSHINAGA, H ;
AMURO, N ;
GOTO, Y ;
OKAZAKI, T .
JOURNAL OF BIOCHEMISTRY, 1994, 115 (02) :194-201
[20]   A PARTIAL DEFICIENCY OF CYTOCHROME-C OXIDASE IN CHRONIC PROGRESSIVE EXTERNAL OPHTHALMOPLEGIA [J].
JOHNSON, MA ;
TURNBULL, DM ;
DICK, DJ ;
SHERRATT, HSA .
JOURNAL OF THE NEUROLOGICAL SCIENCES, 1983, 60 (01) :31-53