• Journal of Microbiology and Biotechnology
• /
• v.22 no.4
• /
• pp.571-573
• /
• 2012

Glyceraldehyde-3-phosphate (G-3-P), the substrate of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), is a key intermediate in several metabolic pathways. Recently, we reported that G-3-P directly inhibits caspase-3 activity in a reversible noncompetitive mode, suggesting the intracellular G-3-P level as a cell fate decision factor. It has been known that apoptotic stimuli induce the generation of NO, and NO S-nitrosylates GAPDH at the catalytic cysteine residue, which confers GAPDH the ability to bind to Siah-1, an E3 ubiquitin ligase. The GAPDH-Siah-1 complex is translocated into the nucleus and subsequently triggers the apoptotic process. Here, we clearly showed that intracellular G-3-P protects GAPDH from S-nitrosylation at above a certain level, and consequently maintains the cell survival. In case G-3-P drops below a certain level as a result of exposure to specific stimuli, G-3-P cannot inhibit S-nitrosylation of GAPDH anymore, and consequently GAPDH translocates with Siah-1 into the nucleus. Based on these results, we suggest that G-3-P functions as a molecule switch between cell survival and apoptosis by regulating S-nitrosylation of GAPDH.

• Asian-Australasian Journal of Animal Sciences
• /
• v.21 no.11
• /
• pp.1673-1679
• /
• 2008

In order to study the biological function of gapdh gene in yak, and prove whether the gapdh gene was a useful intra-reference gene that can be given an important role in molecular biology research of yak, the cDNA sequence encoding glyceraldehyde-3-phosphate dehydrogenase from yak was cloned by the RT-PCR method using gene specific PCR primers. The sequence results indicated that the cloned cDNA fragment (1,008 bp) contained a 1,002 bp open reading frame, encoding 333 amino acids (AAs) with a molecular mass of 35.753 kDa. The deduced amino acids sequence showed a high level of sequence identity to Bos Taurus (99.70%), Xenopus laevis (94.29%), Homo sapiens (97.01%), Mus musculus (97.90%) and Sus scrofa (98.20%). The expression of yak's gapdh gene in heart, spleen, kidney and brain tissues was also detected; the results showed that the gapdh gene was expressed in all these tissues. Further analysis of yak GAPDH amino acid sequence implied that it contained a complete glyceraldehyde-3-phosphate dehydrogenase active site (ASCTTNCL) which ranged from 148 to 155 amino acid residues. It also contained two conserved domains, a NAD binding domain in its N-terminal and a complete catalytic domain of sugar transport in its C-terminal. The phylogenetic analysis showed that yak and Bos taurus were the closest species. The prediction of secondary structures indicated that GAPDH of yak had a similar secondary structure to other isolated GAPDH. The results of this study suggested that the gapdh gene of yak was similar to other species and could be used as the intra-reference to analyze the expression of other genes in yak.

• Fisheries and Aquatic Sciences
• /
• v.11 no.3
• /
• pp.177-181
• /
• 2008

Glyceraldehyde 3-phosphate dehydrogenase(GAPDH) is a key enzyme for carbohydrate metabolism in most living organisms. Recent reports and our own searches of teleost species in publicly available genomic databases have identified at least two distinct GAPDH genes in a given species. The two GAPDH genes are located on the same chromosome in teleosts, whereas they are located on the different chromosomes in mammals. Thus, we reconstructed a phylogenetic tree to better understand the evolutionary history of the GAPDH genes in the vertebrate lineage. Our phylogenetic analysis revealed unambiguously that the two GAPDH genes of teleosts are phylogenetically closely affiliated to one of the cytosolic GAPDH and spermatogenic GAPDH-S of mammals. This indicates that the two paralogous GAPDH genes shared a common ancestor and subsequently underwent a gene duplication event during early vertebrate evolution. However, GAPDH-S of teleosts showed significant differences in the polypeptide residues and tissue distribution of its mRNA transcripts from that of mammals, implying they have undergone a different history of functionalization.

• Journal of fish pathology
• /
• v.28 no.1
• /
• pp.9-15
• /
• 2015

The potential of Streptococcus iniae glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as an antigen for a subunit vaccine was investigated using a zebrafish model. The recombinant S. iniae GAPDH was purified using His-tag column chromatography, and antisera against the recombinant GAPDH (rGAPDH) were produced by intraperitoneal immunization of rats. By immunization with S. iniae rGAPDH, the survival rates of zebrafish against an S. iniae challenge increased, suggesting that GAPDH would be an antigen capable of inducing protective immune responses in fish. Furthermore, we demonstrated using Western blotting, that the antisera against rGAPDH of S. iniae had cross-reactivity with GAPDH from Streptococcus parauberis and Lactococcus garviae, which are also culprits of streptococcosis in cultured fish in Korea. These results suggest that S. iniae GAPDH may be used as an antigen for the development of a subunit vaccine against streptococcosis caused by diverse cocci in cultured fish.

• Journal of Plant Biotechnology
• /
• v.35 no.2
• /
• pp.95-100
• /
• 2008

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a main enzyme in the glycolytic pathway, is involved in cellular energy production and regarded as a housekeeping gene. Previously, cytosolic GAPDH was selected as the most significantly abundant gene in EST library of sweetpotato suspension cells. In this study, a full-length of cDNA clone (IbGAPDH) encoding GAPDH was isolated from suspension-cultured cells of sweetpotato (Ipomoea babatas), and its expression was investigated with a view to understanding the physiological function of GAPDH in relation to environmental stresses. IbGAPDH encoded a 36.9 kDa polypeptide consisting of 337 amino acids. When the deduced amino acid of IbGAPDH was compared with other higher plants, IbGAPDH showed high homology with cytosolic GAPDH. The mRNA level of IbGAPDH significantly increased under environmental stresses, such as $H_2O_2$, MV and cold treatments. Among them, the transcript level of IbGAPDH gene was the highest under cold stress. Further investigation of the transcription level under $10^{\circ}C$ or $15^{\circ}C$ was performed with different tissues of sweetpotato. The transcription of IbGAPDH was increased by cold stress with tissue-specificity, moreover, showed different patterns according to temperature.

• Molecules and Cells
• /
• v.42 no.8
• /
• pp.597-603
• /
• 2019

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a core enzyme of the aerobic glycolytic pathway with versatile functions and is associated with cancer development. Recently, Kornberg et al. published the detailed correlation between GAPDH and di- or monomethyl fumarate (DMF or MMF), which are well-known GAPDH antagonists in the immune system. As an extension, herein, we report the crystal structure of MMF-bound human GAPDH at $2.29{\AA}$. The MMF molecule is covalently linked to the catalytic Cys152 of human GAPDH, and inhibits the catalytic activity of the residue and dramatically reduces the enzymatic activity of GAPDH. Structural comparisons between $NAD^+$-bound GAPDH and MMF-bound GAPDH revealed that the covalently linked MMF can block the binding of the $NAD^+$ cosubstrate due to steric hindrance of the nicotinamide portion of the $NAD^+$ molecule, illuminating the specific mechanism by which MMF inhibits GAPDH. Our data provide insights into GAPDH antagonist development for GAPDH-mediated disease treatment.

• Journal of Microbiology and Biotechnology
• /
• v.23 no.4
• /
• pp.579-585
• /
• 2013

Streptococcus equi ssp. zooepidemicus (Streptococcus zooepidemicus, SEZ) is an important pathogen associated with opportunistic infections of a wide range of species, including pigs and humans. The absence of a suitable vaccine makes it difficult to control SEZ infection. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has been previously identified as an immunogenic protein using immunoproteomic techniques. In the present study, we confirmed that the sequence of GAPDH was highly conserved with other Streptococcus spp. The purified recombinant GAPDH could elicit a significant humoral antibody response in mice and confer significant protection against challenge with a lethal dose of SEZ. GAPDH could adhere to the Hep-2 cells, confirmed by flow cytometry, and inhibit adherence of SEZ to Hep-2 cells in an adherence inhibition assay. In addition, real-time PCR demonstrated that GAPDH was induced in vivo following infection of mice with SEZ. These suggest that GAPDH could play an important role in the pathogenesis of SEZ infection and could be a target for vaccination against SEZ.

• Journal of Life Science
• /
• v.20 no.12
• /
• pp.1743-1749
• /
• 2010

A research group demonstrated that the 37 kDA protein of Edwardsiella tarda, a causing causative agent of edwardsiellosis in fish, exhibited high antigenicity in Japanese flounder. The research group also showed that the N-terminus amino acid sequences of the 37 kDa protein were mapped to the N-terminus of GAPDH (glyceraldehyde-3-phosphate dehydrogenase). Using degenerated primer sets based on the known N-terminus sequence, the corresponding E. tarda DNA was amplified and cloned. The nucleotide sequences of the cloned gene revealed high homology with a bacterial gene for GAPDH, as we was expected. The amino acid sequence of E. tarda GAPDH (etGAPDH) revealed a <70% similarity with GAPDH proteins in other Enterobacteriaceae. With the application of artificial protein overexpression system in Escherichia coli, the recombinant etGAPDH (rGAPDH) was produced and purified. In this study, Using the purified rGAPDH, the etGAPDH specific polyclonal antibody has been was generated using the purified rGAPDHin this study. The immunoblotting analyses demonstrated that the location of the GAPDH protein is located with the association of is associated with the envelops of E. tarda. The rGAPDH was administrated into Japanese flounder via IP route for evaluation of the protective ability. Although the specific antibody titer against etGAPDH was increased about 3-fold after 4 weeks post-vaccination, the survival rates of vaccinated Japanese flounder and the control group with wild type E. tarda was were 12.5% and 0%, respectively. Our results indicated that rGAPDH is immunoreactive antigen but that it will not generate protective immunity in Japanese flounder.

• The Korean Journal of Mycology
• /
• v.51 no.3
• /
• pp.239-243
• /
• 2023

The Golovinomyces biocellatus complex is known to consist of powdery mildew from the Golovinomyces genus, associated with host plants from the Lamiaceae family. Recent molecular phylogenetic analyses have resolved the taxonomic composition of this complex, and Golovinomyces biocellatus sensu stricto is considered to be a pathogen of Glechoma species, globally. However, this paper presents a new finding of Golovinomyces salviae on Glechoma longituba, besides its original host species of Salvia. This information was inferred by molecular phylogenetic analyses from the multi-locus nucleotide sequence dataset of intergeneric spacer (IGS), internal transcribed spacer (ITS), large subunit (LSU) of rDNA, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene. Further, the asexual morphology of this fungus is described and illustrated.

• Molecules and Cells
• /
• v.28 no.6
• /
• pp.559-563
• /
• 2009

Glyceraldehyde-3-phosphate is a key intermediate in several central metabolic pathways of all organisms. Aldolase and glyceraldehyde-3-phosphate dehydrogenase are involved in the production or elimination of glyceraldehyde-3-phosphate during glycolysis or gluconeogenesis, and are differentially expressed under various physiological conditions, including cancer, hypoxia, and apoptosis. In this study, we examine the effects of glyceraldehyde-3-phosphate on cell survival and apoptosis. Overexpression of aldolase protected cells against apoptosis, and addition of glyceraldehyde-3-phosphate to cells delayed apoptosis. Additionally, delayed apoptotic phenomena were observed when glyceraldehyde-3-phosphate was added to a cell-free system, in which artificial apoptotic process was induced by adding dATP and cytochrome c. Surprisingly, glyceraldehyde-3-phosphate directly suppressed caspase-3 activity in a reversible noncompetitive mode, preventing caspase-dependent proteolysis. Based on these results, we suggest that glyceraldehyde-3-phosphate, a key molecule in several central metabolic pathways, functions as a molecule switch between cell survival and apoptosis.