Amino acid lactate dehydrogenase sequence

An example of an enzyme which has different isoenzyme forms is lactate dehydrogenase (LDH) which catalyzes the reversible conversion of pyruvate into lactate in the presence of the coenzyme NADH (see above). LDH is a tetramer of two different types of subunits, called H and M, which have small differences in amino acid sequence. The two subunits can combine randomly with each other, forming five isoenzymes that have the compositions H4, H3M, H2M2, HM3 and M4. The five isoenzymes can be resolved electrophoretically (see Topic B8). M subunits predominate in skeletal muscle and liver, whereas H subunits predominate in the heart. H4 and H3M isoenzymes are found predominantly in the heart and red blood cells H2M2 is found predominantly in the brain and kidney while HM3 and M4 are found predominantly in the liver and skeletal muscle. Thus, the isoenzyme pattern is characteristic of a particular tissue, a factor which is of immense diagnostic importance in medicine. Myocardial infarction, infectious hepatitis and muscle diseases involve cell death of the affected tissue, with release of the cell contents into the blood. As LDH is a soluble, cytosolic protein it is readily released in these conditions. Under normal circumstances there is little LDH in the blood. Therefore the pattern of LDH isoenzymes in the blood is indicative of the tissue that released the isoenzymes and so can be used to diagnose a condition, such as a myocardial infarction, and to monitor the progress of treatment. 

Predict its most likely protein translation by submitting the DNA sequence along with the following amino acid sequences of human lactate dehydrogenase isozymes to Procustes analyses. 

The amino acid sequences of corresponding regions from dogfish M4 and Bacillus stearothermophilus lactate dehydrogenases are shown below ... 

Fields, P.A., and G.N. Somero (1997). Amino acid sequence differences cannot fully explain interspecific variation in thermal sensitivities of gobiid fish A4-lactate dehydrogenases (A4-LDHs). J. Exp. Biol. 200 1839-1850. 

Labeyrie et al. (41) isolated a trypsin fragment of 11 kDa from S. cerevisiae flavocytochrome 62 that contained heme but was devoid of flavin and had no lactate dehydrogenase activity. The fragment, which was referred to as cytochrome 62 core, was found to have spectral properties very like those of microsomal cytochrome 65 (41). This similarity with cytochrome 65 is borne out by comparisons of amino acid sequence (42-44). The sequence similarity extends to related heme domains of sulfite oxidase (45, 46) and assimilatory nitrate reductase (47). The existence of a protein family with a common cytochrome 65 fold was suggested by Guiard and Lederer (48) and this is supported by the close similarity between the three-dimensional structures of microsomal cytochrome 65 (49) and the cytochrome domain of flavocytochrome 62 (23-25). 

Limited amino acid sequence information has shown that long-chain a-hydroxyacid oxidase from rat kidney is also related to these FMN-containing oxidoreductases (55). It is likely that several further members of this family remain to be identified. The flavodehydrogenase domain shows no sequence similarity to the lactate dehydrogenase from bacteria and higher eukaryotes that utilize NAD as a substrate. Yeasts lack such an enzyme and the substrate specificity of flavocytochrome 62 has presumably evolved independently of the NAD-linked dehydrogenases. 

Lactate dehydrogenase was the first enzyme that established the structural basis for the existence of isoenzymes (different forms of an enzyme resulting from variations in amino acid sequence). Most tissues contain five isoenzymes of lactate dehydrogenase that can be resolved electrophoretically. 

Lactate dehydrogenase (LDH) is a tetrameric protein consisting of two types of subunits, called M and H, which have small differences in amino acid sequence. Different molecular forms of an enzyme are called isoenzymes or isozymes. 

---