Isolation and Characterization of cDNA Clones Encoding Human Liver Glutamate Dehydrogenase: Evidence for a Small Gene Family

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 85; no. 10; pp. 3494 - 3498
• Main Authors: Mavrothalassitis, George, Tzimagiorgis, George, Mitsialis, Alex, Zannis, Vassilis, Plaitakis, Andreas, Papamatheakis, Joseph, Moschonas, Nicholas
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 01.05.1988; National Acad Sciences
• Subjects: 550201 - Biochemistry- Tracer Techniques; Adult; AMINE OXIDASES; AMINO ACID SEQUENCE; ANIMAL CELLS; ANIMALS; Base Sequence; BASIC BIOLOGICAL SCIENCES; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; Biological and medical sciences; Biotechnology; BODY; Cloning, Molecular; Complementary DNA; CONNECTIVE TISSUE CELLS; DAYS LIVING RADIOISOTOPES; DIGESTIVE SYSTEM; DNA; DNA - genetics; DNA - isolation & purification; DNA probes; ENZYMES; Fetus; FIBROBLASTS; Fundamental and applied biological sciences. Psychology; Generally accepted auditing standards; Genes; Genetic engineering; Genetic technics; Gin; GLANDS; Glutamate Dehydrogenase - genetics; Humans; HYBRIDIZATION; ISOTOPES; LIGHT NUCLEI; LIVER; Liver - enzymology; Macaca fascicularis; MAMMALS; MAN; Methods. Procedures. Technologies; Molecular cloning; Molecular Sequence Data; MOLECULAR STRUCTURE; NUCLEI; Nucleic Acid Hybridization; NUCLEIC ACIDS; ODD-ODD NUCLEI; Oligonucleotide probes; OLIGONUCLEOTIDES; Organ Specificity; ORGANIC COMPOUNDS; ORGANS; OXIDOREDUCTASES; PHOSPHORUS 32; PHOSPHORUS ISOTOPES; PRIMATES; Rabbits; RADIOISOTOPES; RECOMBINANT DNA; RNA; SOMATIC CELLS; Species Specificity; VERTEBRATES; Characterization; Glutamate dehydrogenase; Human; Complementary DNA; Gene; Liver; Isolation; Multigene family; Molecular cloning
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.85.10.3494

We have isolated a series of human liver cDNA clones encoding glutamate dehydrogenase. The cDNA-derived protein sequence specifies a single 558-amino acid long polypeptide including a cleavable signal sequence of 53 amino acids. Blotting analysis of RNA from human, monkey, and rabbit showed that glutamate dehydrogenase mRNA is present in various amounts in all tissues tested. Glutamate dehydrogenase mRNAs are of four sizes and are found in different ratios in different tissues; the predominant ones are ≈ 3.5 and ≈ 2.9 kilobases. Blot hybridization of human genomic DNA to nonoverlapping cDNA fragments revealed multiple bands, many of which hybridize with two or more probes in a manner inconsistent with the existence of a single GLUD gene. Moreover, two separate 36-base synthetic oligonucleotides corresponding to the coding region hybridize to multiple genomic fragments, confirming the existence of more than one GLUD-related gene in human.