TY - JOUR
T1 - Use of denaturing gradient gel electrophoresis for detection of mutation and prospective diagnosis in late onset ornithine transcarbamylase deficiency
AU - Finkelstein, Janice E.
AU - Francomano, Clair A.
AU - Brusilow, Saul W.
AU - Traystman, Monica D.
N1 - Funding Information:
The authors thank Dr. Haig Kazazian for his expert advice, Dr. Mendel Tuchman for performing enzyme analysis on liver tissue, Dr. Beth Dombroski for her technical advice and assistance with sequencing, and Ellen Gordes and Evelyn Bull for their expert technical assistance in performing the biochemical assays. J.E.F. is supported by a National Institute of Health Medical Genetics Fellowship Training Grant (GM 07471) and a grant from the Stetler Research Fund for Women Physicians. This work was further supported by NIH Grant ROl HD11134 and grants from the Kettering Family Foundation and the TA and MA O’Malley Foundation to S.B., NIH Grant Kll AM01361 to C.A.F., and a Judith Graham Pool Post-graduate Research Fellowship awarded by the National Hemophilia Foundation to M.D.T. Clinical studies were performed in the Pediatric Clinical Research Unit of the Johns Hopkins Hospital and supported by NIH Grant MO1 RR00052.
PY - 1990/6
Y1 - 1990/6
N2 - Ornithine transcarbamylase (ornithine carbamoyl-transferase, EC 2.1.3.3) deficiency is an X-linked inborn error of metabolism with considerable phenotypic variability in affected males. Using a combination of the polymerase chain reaction and denaturing gradient gel electrophoresis (DGGE), we defined a mutation in a family in whom affected males have significant residual enzyme activity. A C → T change in the first nucleotide of codon 277 resulted in the substitution of a tryptophan for an arginine at amino acid 245 of the mature protein. This change appears to represent a deleterious mutation rather than a polymorphism on the basis of several factors: the change occurs at a highly conserved arginine residue, significant size and change differences exist between arginine and tryptophan, and this change was not seen on DGGE screening of 26 unrelated individuals representing 43 chromosomes. Diagnosis of an at-risk male newborn in this family was performed using direct mutational analysis. In families with partial enzyme deficiencies in whom biochemical data may be difficult to evaluate, direct detection of mutations at the OTC locus permits definitive diagnosis. This represents the first description of a mutation in late onset OTC deficiency and demonstrates direct mutational analysis by DGGE for prospective diagnosis in a genetic disorder.
AB - Ornithine transcarbamylase (ornithine carbamoyl-transferase, EC 2.1.3.3) deficiency is an X-linked inborn error of metabolism with considerable phenotypic variability in affected males. Using a combination of the polymerase chain reaction and denaturing gradient gel electrophoresis (DGGE), we defined a mutation in a family in whom affected males have significant residual enzyme activity. A C → T change in the first nucleotide of codon 277 resulted in the substitution of a tryptophan for an arginine at amino acid 245 of the mature protein. This change appears to represent a deleterious mutation rather than a polymorphism on the basis of several factors: the change occurs at a highly conserved arginine residue, significant size and change differences exist between arginine and tryptophan, and this change was not seen on DGGE screening of 26 unrelated individuals representing 43 chromosomes. Diagnosis of an at-risk male newborn in this family was performed using direct mutational analysis. In families with partial enzyme deficiencies in whom biochemical data may be difficult to evaluate, direct detection of mutations at the OTC locus permits definitive diagnosis. This represents the first description of a mutation in late onset OTC deficiency and demonstrates direct mutational analysis by DGGE for prospective diagnosis in a genetic disorder.
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U2 - 10.1016/0888-7543(90)90537-5
DO - 10.1016/0888-7543(90)90537-5
M3 - Article
C2 - 2347583
AN - SCOPUS:0025281536
SN - 0888-7543
VL - 7
SP - 167
EP - 172
JO - Genomics
JF - Genomics
IS - 2
ER -