Molecular characterization and expression pattern of the equine lactate dehydrogenase A and B genes.
Abstract: The species-specific properties of LDH isozymes are essentially determined by M (muscle) and H (heart) subunit proteins encoded by the LDHA and LDHB genes, respectively. In the present study, we molecularly characterized the full-length equine lactate dehydrogenase A (eLDHA) and B (eLDHB) cDNAs. The eLDHA cDNA consisted of a 999-bp open reading frame (ORF), while the eLDHB and newly acquired bat LDHB consisted of a 1002-bp ORF, which is 3 bp shorter than the LDHB ORF of other registered mammals. The alignment of amino acid sequences showed that eLDHA acquired positively charged His 88 and 226, and eLDHB lost negatively charged Glu 14, as compared to the highly conserved residues at these positions in the corresponding amino acid sequences of other mammals. These alterations were identified in six equine species by genomic DNA analysis. A comparison of the equine and human 3D structures revealed that the substituted His 88 and 226 of the eLDHA monomer and the deleted Glu 14 of the eLDHB monomer altered the surface charge of equine LDH tetramers and that these three residues were located in important regions affecting the catalytic kinetics. Also, RT-PCR amplification of the three myosin heavy chain isoforms corroborated that the cervical muscle as postural muscle of the thoroughbred horse was composed of more oxidative myofibers than the dynamic muscle. Based on this property, the mRNA expression patterns of eLDHA, eLDHB, and eGAPDH in various tissues were analyzed by using real-time PCR. The expression levels of these three genes in the cervical muscle were not always relatively higher than in the brain or heart.
Publication Date: 2009-07-30 PubMed ID: 19647052DOI: 10.1016/j.gene.2009.07.017Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research investigates the molecular characteristics and expression patterns of two genes (LDHA and LDHB) encoding lactate dehydrogenase (LDH) enzymes in horses. In-depth examination revealed that expression levels of these genes vary across different tissues within the horse’s body.
Molecular Characterization of eLDHA and eLDHB
- The researchers identified the full-length equine lactate dehydrogenase A (eLDHA) and B (eLDHB) cDNAs, which are the genetic blueprints for creating LDH proteins. LDH A and B carry out crucial biochemical processes in horses and other mammals.
- They noticed that the eLDHA cDNA had an open reading frame (ORF) of 999 base pairs, while eLDHB had an ORF of 1002 base pairs, slightly shorter than that observed in other mammals.
Distinct Amino Acid Sequences
- The amino acid sequences of eLDHA and eLDHB showed distinct differences from their counterparts in other mammals. The eLDHA sequence had additional positively charged Histidine residues at positions 88 and 226. The eLDHB sequence was missing a negatively charged Glutamate or Glu 14, which is typically conserved in other mammals.
- These changes in amino acid sequences were confirmed in six different horse species via genomic DNA analysis.
Structural and Functional Implications
- Comparing the 3D structures of the human and horse LDH proteins revealed that the substitution and deletion of certain amino acids in the horse proteins altered surface charges. These changes may impact the functionality and catalytic kinetics of the proteins.
- The residues (His 88 and 226 in eLDHA, Glu14 in eLDHB) were shown to be placed in significant regions that affect catalytic kinetics or the speed and efficiency of the biochemical reactions that these enzymes facilitate.
Expression Patterns of eLDHA and eLDHB
- Using Real-Time Polymerase Chain Reaction (PCR), the researchers looked at the expression of thse LDH genes in different tissues. They discovered that the expression levels were not consistently higher in the horse’s cervical muscle (part of the neck) compared to the brain or heart, despite the fact that cervical muscles in thoroughbred horses typically have more oxidative myofibers, which use more oxygen and are associated with endurance activities.
Cite This Article
APA
Echigoya Y, Sato T, Itou T, Endo H, Sakai T.
(2009).
Molecular characterization and expression pattern of the equine lactate dehydrogenase A and B genes.
Gene, 447(1), 40-50.
https://doi.org/10.1016/j.gene.2009.07.017 Publication
Researcher Affiliations
- Nihon University Veterinary Research Center, 1866 Kameino, Fujisawa, Kanagawa 252-8510, Japan.
MeSH Terms
- Amino Acid Sequence
- Animals
- DNA, Complementary / chemistry
- Exons
- Gene Expression
- Horses / genetics
- L-Lactate Dehydrogenase / chemistry
- L-Lactate Dehydrogenase / genetics
- Models, Molecular
- Molecular Sequence Data
- Open Reading Frames
- Phylogeny
- Protein Conformation
- RNA, Messenger / metabolism
- Sequence Alignment
Citations
This article has been cited 6 times.- Wu J, Chen Y, Lin Y, Lan F, Cui Z. Cancer-testis antigen lactate dehydrogenase C4 as a novel biomarker of male infertility and cancer. Front Oncol 2022;12:936767.
- Yao H, Yang F, Li Y. Natural products targeting human lactate dehydrogenases for cancer therapy: A mini review. Front Chem 2022;10:1013670.
- Xu Q, Li X, Ma L, Loor JJ, Coleman DN, Jia H, Liu G, Xu C, Wang Y, Li X. Adipose tissue proteomic analysis in ketotic or healthy Holstein cows in early lactation1. J Anim Sci 2019 Jul 2;97(7):2837-2849.
- Chen IH, Wang JH, Chou SJ, Wu YH, Li TH, Leu MY, Chang WB, Yang WC. Selection of reference genes for RT-qPCR studies in blood of beluga whales (Delphinapterus leucas). PeerJ 2016;4:e1810.
- Chen IH, Chou LS, Chou SJ, Wang JH, Stott J, Blanchard M, Jen IF, Yang WC. Selection of suitable reference genes for normalization of quantitative RT-PCR in peripheral blood samples of bottlenose dolphins (Tursiops truncatus). Sci Rep 2015 Oct 21;5:15425.
- Echigoya Y, Okabe H, Itou T, Endo H, Sakai T. Molecular characterization of glycogen synthase 1 and its tissue expression profile with type II hexokinase and muscle-type phosphofructokinase in horses. Mol Biol Rep 2011 Jan;38(1):461-9.
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