GALK1

Case study 1: Martyn A Sharpe, 2021

Glioblastoma (GBM) can use metabolic fuels other than glucose (Glc). The ability of GBM to use galactose (Gal) as a fuel via the Leloir pathway is investigated. Gene transcript data were accessed, growth studies were performed and Gal metabolism was examined using proton, carbon and phosphorous NMR spectroscopy.

Data analysis from published databases revealed that elevated levels of mRNA transcripts of SLC2A3 (Glut3), SLC2A14 (Glut14) and key Leloir pathway enzymes correlate with poor patient outcomes. The 13C NMR-based metabolic flux analysis demonstrated a fully functional Leloir pathway and elevated pentose phosphate pathway activity for efficient Gal metabolism in GBM cells. In all, the expression of Glut3 and/or Glut14 together with the enzymes of the Leloir pathway allows GBM to transport and metabolize Gal at physiological glucose concentrations, providing GBM cells with an alternate energy source.

Fig1. Correlation between the expression levels of Gal import genes (SLC2A3, SLC2A14, GALM, GALK1, GALE, PGM1/2 and UGP2).

Fig2. Five primary cell cultures were incubated for 24 h in 10 μM CDP36, a GALK1-specific inhibitor, and underwent a loss in viability only in Gal-containing, and not Glc-containing.

Case study 2: James M Termini, 2017

Here the researchers describe derivatives of the HEK293T cell line that are defective in their ability to generate mucin-type O-linked glycosylation. Using CRISPR/Cas9 and a single-cell GFP-sorting procedure, the UDP-galactose-4-epimerase (GALE), galactokinase 1 (GALK1), and galactokinase 2 (GALK2) genes were knocked out individually and in combinations with greater than 90% of recovered clones having the desired mutations. GALK1 and GALK2 are key enzymes of the galactose/GalNAc salvage pathways. Mass spectrometry was performed on whole cell lysate of the knockout cell lines to verify the glycosylation phenotype. Mass spectrometry analysis revealed that the disruption of GALE, GALK1, and GALE+GALK2 had little effect on the N-glycome. But when GALE was knocked out in tandem with GALK1, N-glycans were exclusively of the high mannose type. Due to the well-characterized nature of these five knockout cell lines, they will likely prove useful for a wide variety of applications.

Fig3. Validation of GALK1 and GALE+GALK1 deficient knockout cell lines.

Fig4. HEK293T cells, GALE KO (clone 4), GALK1 KO (clone 10), GALE+GALK1 KO (clone 12), GALK2 KO (clone 7), and GALE+GALK2 KO (clone 7) were analyzed to determine if disrupting key enzymes of the Leloir pathway impacted cell growth.

Fig1. Ilustration of the metabolism of galactose through the Leloir pathway. (Martyn A Sharpe, 2021)

Fig2. Leloir pathway of galactose metabolism. Illustrated are the 2 different pathways in which galactose and GalNAc can be salvaged or synthesized for use in glycosylation. (James M Termini, 2017)

GALK1 involved in several pathways and played different roles in them. We selected most pathways GALK1 participated on our site, such as Galactose metabolism, Amino sugar and nucleotide sugar metabolism, Metabolic pathways, which may be useful for your reference. Also, other proteins which involved in the same pathway with GALK1 were listed below. Creative BioMart supplied nearly all the proteins listed, you can search them on our site.

GALK1 has several biochemical functions, for example, ATP binding, galactokinase activity, galactose binding. Some of the functions are cooperated with other proteins, some of the functions could acted by GALK1 itself. We selected most functions GALK1 had, and list some proteins which have the same functions with GALK1. You can find most of the proteins on our site.

GALK1 has direct interactions with proteins and molecules. Those interactions were detected by several methods such as yeast two hybrid, co-IP, pull-down and so on. We selected proteins and molecules interacted with GALK1 here. Most of them are supplied by our site. Hope this information will be useful for your research of GALK1.

OTUD4; DMWD; 15-deoxy-delta(12,14; ESR1; VCAM1; CD9; AGO4