Summary
We have previously detected a single base substitution of G by A at the Arg codon CGC in exon 4 of the mutant lactate dehydrogenase (LDH) gene, an unstable LDH-B variant (case 1). Here, we use the polymerase chain reaction (PCR) to amplify genomic DNA of two cases (the original case 1 and a new patient, case 2). We were able to confirm that case 1 is homozygous for the mutation, causing a replacement of the conserved Arg by His at residue 173. The resulting LDH-B variant subunit is unstable in vivo. Whereas the mutation in exon 4 was not observed in case 2, a different single base substitution of A by C was detected at the Ser codon AGT in exon 3. This mutation causes a replacement of the conserved Ser by Arg at residue 131. Genomic analysis of the family of case 2 by mismatched PCR showed that the missense mutation was consistent with their biochemical phenotypes. The replacement results in a conformational change of the residues near the Ser, probably because the side chain of Arg is much more bulky than that of Ser. The change may affect the arrangement of the cofactor binding site and result in the loss of enzyme activity. The experimental observations are consistent with computer graphics analyses.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Boone C, Chen TR, Ruddle FH (1972) Assignment of three human genes to chromosomes (LDH-A to 11, TK to 17 and IDH to 10) and evidence for translocation between human and mouse chromosomes in somatic cells hybrids. Proc Natl Acad Sci USA 69:510–514
Chung FZ, Tsujibo H, Bhattacharyya U, Sharief FS, Li SS-L (1985) Genomic organization of human lactate dehydrogenase A gene Biochem J 231:537–541
Clarf AR, Wigly DB, Barstow DA, William NC, Atkinson T, Holbrook JJ (1987) A single amino acid substitution deregulates a bacterial lactate dehydrogenase and stabilizes its tetrameric structure. Biochim Biophys Acta 931:72–80
Eventoff W, Rossmann MG, Taylor SS, Torff HJ, Meyer H, Keil W, Kiltz HH (1977) Structural adaptations of lactate dehydrogenase isozymes. Proc Natl Acad Sci USA 74:2677–2681
Franke U, Busby N (1975) Assignment of human genes for lactate dehydrogenase A and thymidine kinase to specific chromosome regions. Cytogenet Cell Genet 14:313–319
Grau UM, Trommer WE, Rossmann MG (1981) Structure of the activity ternary complex of pig heart lactate dehydrogenase with S-lac-NAD at 2.7 Å resolution. J Mol Biol 151:289–307
Gyllenstein UB, Erlich HA (1988) Generation of single stranded DNA by the polymerase chain reaction and its application to direct sequencing of the HAL-DQA locus. Proc Natl Acad Sci USA 85:7652–7656
Hiraoka BY, Sharief FS, Yang YW, Li WH, Li SS-L (1990) The cDNA and protein sequences of mouse lactate dehydrogenase B: molecular evolution of vertebrate LDH genes A(muscle), B(heart) and C(testis). Eur J Biochem 189:215–220
Houki N, Matsushima Y, Kitamura M, Tsukada T, Nishina T, Nakayama T (1986) A case of deficiency of lactate dehydrogenase H-subunit. Jpn J Clin Chem 15:85–90
Joukyuu R, Mizuno S, Amakawa T, Tsukada T, Nishina T, Kitamura M (1989) Hereditary complete deficiency of lactate dehydrogenase H-subunit. Clin Chem 35:687–690
Kamata M, Taninaka S, Inaba R, Fujita K, Sakurabayashi I, Tsukada T, Kitamura M (1989) A case with lactate dehydrogenase H subunit variant. Seibutu Buturi Kagaku 33:243
Kanno T, Sudo K, Maekawa M, Nishimura Y, Ukita M, Fukutake K (1988) Lactate dehydrogenase M-subunit deficiency: a new type of hereditary exertional myopathy. Clin Chim Acta 173:89–98
Kao RT, Wu KC, Law ML, Hartz JA, Lau Y-F (1988) Assignment of human gene encoding testis-specific lactate dehydrogenase C to chromosome 11, region p14.3–15.5. Somat Mol Cell Genet 14:515–518
Kitamura M, Iijima N, Hashimoto F, Hiratsuka A (1971) Hereditary deficiency of subunit H of lactate dehydrogenase. Clin Chim Acta 34:419–423
Kumar R, Dunn LL (1989) Designed diagnostic restriction fragment length polymorphisms for the detection of point mutations in ras oncogenes. Oncogene Res 4:235–241
Kunkel LM, Smith KD, Boyer SH, Borgaonkar DS, Wachtel SS, Miller OJ, Breg WR (1977) Analysis of human γ-chromosome-specific reiterated DNA in chromosome variants. Proc Natl Acad Sci USA 74:1245–1249
Li SS-L, Fitch WM, Pan Y-CE, Sharief FS (1983) Evolutionary relationship of vertebrate lactate dehydrogenase isozymes A4 (muscle), B4 (heart), and C4 (testis) J Biol Chem 258:7029–7032
Li SS-L, Tiano HF, Fukasawa KM, Yagi K, Simizu M, Sharief FS, Nakashima Y, Pan Y-CE (1985) Protein structure and gene organization of mouse lactate dehydrogenase A isozyme. Eur J Biochem 149:215–225
Maekawa M, Kanda S, Sudo K, Kanno T (1984) Estimation of the gene frequency of lactate dehydrogenase subunit deficiencies. Am J Hum Genet 36:1204–1214
Mayeda K, Weiss L, Linahl R, Dully M (1974) Localization of the human lactate dehydrogenase B gene on the short arm of chromosome 12. Am J Hum Genet 26:59–64
Mohrenweisser HW, Novotny JE (1982) An enzymatically inactive variant of human lactate dehydrogenase LDH-B Gua-1. Biochem Biophys Acta 702:90–98
Pan Y-CE, Sharief FS, Okabe M, Huang S, Li SS-L (1983) Amino acid sequence studies on lactate dehydrogenase C4 isozymes from mouse and rat testes. J Biol Chem 258:7005–7016
Rossmann MG, Adams MJ, Buehner M, Ford GC, Hackert ML, Liljas A, Rao ST, Banaszak LJ, Hill E, Tsernoglou D, Webb L (1973) Molecular symmetry axes and subunit interfaces in certain dehydrogenases. J Mol Biol 76:533–537
Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA (1988) Primer-directed enzymatic amplification of DNA with a theromostable DNA polymerase. Science 239:487–491
Sakai I, Sharief FS, Pan Y-CE, Li SS-L (1987a) The cDNA and protein sequences of human lactate dehydrogenase-B. Biochem J 248:933–936
Sakai I, Sharief FS, Li SS-L (1987b) Molecular and nucleotide sequence of the cDNA for sperm-specific lactate dehydrogenaseC from mouse. Biochem J 242:619–622
Sudo K, Maekawa M, Ikawa S, Machida K, Kitamura M, Li SS-L (1990) A missense mutation found in human lactate dehydrogenase-B(H) variant gene. Biochem Biophys Res Commun 168:672–676
Takano T, Li SS-L (1989a) Structure of human lactate dehydrogenase-B gene. Biochem J 257:921–924
Takano T, Li SS-L (1989b) Human testicular lactate dehydrogenase-C gene is interrupted by six introns at positions homologous to those of LDH-A (muscle) and LDH-B (heart) genes. Biochem Biophys Res Commun 159:579–583
Taylor SS (1977) Amino acid sequence of dogfish muscle lactate dehydrogenase. J Biol Chem 252:1799–1806
Tomonaga A, Kasai K, Omokawa H, Matsuura Y (1985) A new computer system for the assistance of molecular design. In: Proceedings of 13th Symposium of Chemical Information and Computer Science, Kanazawa, Japan, pp 134–137
Tsujibou H, Tiano HF, Li SS-L (1985) Nucleotide sequences of the cDNA and an intronless pseudogene for human lactate dehydrogenase-A isozymes. Eur J Biochem 147:9–15
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Sudo, K., Maekawa, M., Tomonaga, A. et al. Molecular characterization of genetic mutations in human lactate dehydrogenase (LDH) B (H) variant. Hum Genet 89, 158–162 (1992). https://doi.org/10.1007/BF00217116
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00217116