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Proteintech slc35a1

Effect of p.Thr45Ala on <t>SLC35A1</t> expression and transporter activity. (a) Illustration of the cellular route of Neu5Ac. CMP-Neu5Ac is generated by the bifunctional UDP- N -acetylglucosamine 2-epimerase/ N -acetylmannosamine kinase (GNE) converting UDP-GlcNAc to ManNAc and then to ManNAc-6-phosphate. Next, sialic acid synthase (NANS) and N -acetylneuraminate-9-phosphatase (NANP) convert ManNAc-6-phosphate to Neu5Ac via Neu5Ac-9-phosphate. The final step of CMP-Neu5Ac synthesis occurs in the nucleus, where Neu5Ac is linked to the nucleotide CMP by cytidine monophosphate N -acetylneuraminic acid synthetase (CMAS). CMP-Neu5Ac can also be generated through an alternative synthesis pathway, where sialic acid molecules that have been cleaved from glycoconjugates by the lysosomal neuraminidases NEU1-4 are recycled and subsequently reactivated with CMP in the nucleus. (b) Top: variant c.133A>G of Patient 6 localizes to Exon 2 of the SLC35A1 gene. Bottom: electropherogram (Sanger sequencing) of Exon 2 showing homozygosity for variant c.133A>G; p.Thr45Ala in fibroblasts of Patient 6. (c) Transcript and protein expression determined by qPCR and SLC35A1-ELISA revealed a normal mRNA level but significantly reduced SLC35A1 protein level in the patient's cells. (d) Measurement of the import activity of radioactively labelled sugar substrates into Golgi vesicles isolated from control and patient fibroblasts and normalisation of the CMP-[ 14 C]-Neu5Ac import to the import of control sugars GDP-[ 14 C]-fucose and UDP-[ 14 C]-galactose. (e) Protein structure analysis. Left: visualization was performed by PROTTER (Version 1.0; http://wlab.ethz.ch/protter/start/ ). Highlighted in different colours are variants of the six known SLC35A1-CDG patients and of amino acid residues which are important for substrate binding and activity. Threonine 45 (blue circle) of Patient 6 is predicted to be located at the border of the first luminal loop to the second transmembrane domain. Right: due to the p.Thr45Ala variant (Patient 6), an interhelical H-bond is predicted to get lost. Simulation based on the murine Slc35a1 structure generated by DynaMut (PDB: 6XBO). (f) Cycloheximide (CHX) treatment of SLC35A1 K.o. HEK293 cells expressing the wild-type (WT) or the mutated (p.Thr45Ala) SLC35A1-HA protein revealed a shorter half-life of the mutated SLC35A1. The relative amount of SLC35A1-HA was determined by quantification of the Western blot signals where the start of treatment was set to 100%. The regression lines were used to calculate the protein half-life ( y = 50%).

Slc35a1, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/cmp/SLC35A1+Antibody/pmc12226171-86-5-6
Average 93 stars, based on 11 article reviews

slc35a1 - by Bioz Stars, 2026-11

93/100 stars

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1) Product Images from "A Novel Missense Variant in Ultrarare SLC35A1-CDG Alters Cellular Glycosylation, Lipid, and Energy Metabolism Without Affecting CDG Serum Markers"

Article Title: A Novel Missense Variant in Ultrarare SLC35A1-CDG Alters Cellular Glycosylation, Lipid, and Energy Metabolism Without Affecting CDG Serum Markers

Journal: Human Mutation

doi: 10.1155/humu/6290620

Effect of p.Thr45Ala on SLC35A1 expression and transporter activity. (a) Illustration of the cellular route of Neu5Ac. CMP-Neu5Ac is generated by the bifunctional UDP- N -acetylglucosamine 2-epimerase/ N -acetylmannosamine kinase (GNE) converting UDP-GlcNAc to ManNAc and then to ManNAc-6-phosphate. Next, sialic acid synthase (NANS) and N -acetylneuraminate-9-phosphatase (NANP) convert ManNAc-6-phosphate to Neu5Ac via Neu5Ac-9-phosphate. The final step of CMP-Neu5Ac synthesis occurs in the nucleus, where Neu5Ac is linked to the nucleotide CMP by cytidine monophosphate N -acetylneuraminic acid synthetase (CMAS). CMP-Neu5Ac can also be generated through an alternative synthesis pathway, where sialic acid molecules that have been cleaved from glycoconjugates by the lysosomal neuraminidases NEU1-4 are recycled and subsequently reactivated with CMP in the nucleus. (b) Top: variant c.133A>G of Patient 6 localizes to Exon 2 of the SLC35A1 gene. Bottom: electropherogram (Sanger sequencing) of Exon 2 showing homozygosity for variant c.133A>G; p.Thr45Ala in fibroblasts of Patient 6. (c) Transcript and protein expression determined by qPCR and SLC35A1-ELISA revealed a normal mRNA level but significantly reduced SLC35A1 protein level in the patient's cells. (d) Measurement of the import activity of radioactively labelled sugar substrates into Golgi vesicles isolated from control and patient fibroblasts and normalisation of the CMP-[ 14 C]-Neu5Ac import to the import of control sugars GDP-[ 14 C]-fucose and UDP-[ 14 C]-galactose. (e) Protein structure analysis. Left: visualization was performed by PROTTER (Version 1.0; http://wlab.ethz.ch/protter/start/ ). Highlighted in different colours are variants of the six known SLC35A1-CDG patients and of amino acid residues which are important for substrate binding and activity. Threonine 45 (blue circle) of Patient 6 is predicted to be located at the border of the first luminal loop to the second transmembrane domain. Right: due to the p.Thr45Ala variant (Patient 6), an interhelical H-bond is predicted to get lost. Simulation based on the murine Slc35a1 structure generated by DynaMut (PDB: 6XBO). (f) Cycloheximide (CHX) treatment of SLC35A1 K.o. HEK293 cells expressing the wild-type (WT) or the mutated (p.Thr45Ala) SLC35A1-HA protein revealed a shorter half-life of the mutated SLC35A1. The relative amount of SLC35A1-HA was determined by quantification of the Western blot signals where the start of treatment was set to 100%. The regression lines were used to calculate the protein half-life ( y = 50%).

Figure Legend Snippet: Effect of p.Thr45Ala on SLC35A1 expression and transporter activity. (a) Illustration of the cellular route of Neu5Ac. CMP-Neu5Ac is generated by the bifunctional UDP- N -acetylglucosamine 2-epimerase/ N -acetylmannosamine kinase (GNE) converting UDP-GlcNAc to ManNAc and then to ManNAc-6-phosphate. Next, sialic acid synthase (NANS) and N -acetylneuraminate-9-phosphatase (NANP) convert ManNAc-6-phosphate to Neu5Ac via Neu5Ac-9-phosphate. The final step of CMP-Neu5Ac synthesis occurs in the nucleus, where Neu5Ac is linked to the nucleotide CMP by cytidine monophosphate N -acetylneuraminic acid synthetase (CMAS). CMP-Neu5Ac can also be generated through an alternative synthesis pathway, where sialic acid molecules that have been cleaved from glycoconjugates by the lysosomal neuraminidases NEU1-4 are recycled and subsequently reactivated with CMP in the nucleus. (b) Top: variant c.133A>G of Patient 6 localizes to Exon 2 of the SLC35A1 gene. Bottom: electropherogram (Sanger sequencing) of Exon 2 showing homozygosity for variant c.133A>G; p.Thr45Ala in fibroblasts of Patient 6. (c) Transcript and protein expression determined by qPCR and SLC35A1-ELISA revealed a normal mRNA level but significantly reduced SLC35A1 protein level in the patient's cells. (d) Measurement of the import activity of radioactively labelled sugar substrates into Golgi vesicles isolated from control and patient fibroblasts and normalisation of the CMP-[ 14 C]-Neu5Ac import to the import of control sugars GDP-[ 14 C]-fucose and UDP-[ 14 C]-galactose. (e) Protein structure analysis. Left: visualization was performed by PROTTER (Version 1.0; http://wlab.ethz.ch/protter/start/ ). Highlighted in different colours are variants of the six known SLC35A1-CDG patients and of amino acid residues which are important for substrate binding and activity. Threonine 45 (blue circle) of Patient 6 is predicted to be located at the border of the first luminal loop to the second transmembrane domain. Right: due to the p.Thr45Ala variant (Patient 6), an interhelical H-bond is predicted to get lost. Simulation based on the murine Slc35a1 structure generated by DynaMut (PDB: 6XBO). (f) Cycloheximide (CHX) treatment of SLC35A1 K.o. HEK293 cells expressing the wild-type (WT) or the mutated (p.Thr45Ala) SLC35A1-HA protein revealed a shorter half-life of the mutated SLC35A1. The relative amount of SLC35A1-HA was determined by quantification of the Western blot signals where the start of treatment was set to 100%. The regression lines were used to calculate the protein half-life ( y = 50%).

Techniques Used: Expressing, Activity Assay, Generated, Variant Assay, Sequencing, Enzyme-linked Immunosorbent Assay, Isolation, Control, Binding Assay, Western Blot

Effect of SLC35A1 deficiency on glycosylation in fibroblasts. (a) Lectin binding studies showed a significant increase of nonsialylated structures (RCAI) and a significant decrease in α -2,3 sialylated glycans (MAL-I) in the patient, whereas the amount of α -2,6 sialylated residues (SNA) in SLC35A1-CDG was comparable to controls. (b) Expression of ICAM1 and GP130 is significantly reduced in the patient's cells (Pat.) compared to controls (Ctrl.). For quantification, the fully glycosylated protein forms (glyco., black arrows) were compared to the hypoglycosylated forms (nonglyco, grey arrows). (c) LC-MS analysis of N-glycans in fibroblasts revealed a deviating amount of individual N-glycans in the patient's cells (indicated by black boxes and arrows).

Figure Legend Snippet: Effect of SLC35A1 deficiency on glycosylation in fibroblasts. (a) Lectin binding studies showed a significant increase of nonsialylated structures (RCAI) and a significant decrease in α -2,3 sialylated glycans (MAL-I) in the patient, whereas the amount of α -2,6 sialylated residues (SNA) in SLC35A1-CDG was comparable to controls. (b) Expression of ICAM1 and GP130 is significantly reduced in the patient's cells (Pat.) compared to controls (Ctrl.). For quantification, the fully glycosylated protein forms (glyco., black arrows) were compared to the hypoglycosylated forms (nonglyco, grey arrows). (c) LC-MS analysis of N-glycans in fibroblasts revealed a deviating amount of individual N-glycans in the patient's cells (indicated by black boxes and arrows).

Techniques Used: Glycoproteomics, Binding Assay, Expressing, Liquid Chromatography with Mass Spectroscopy

Metabolic measurements and GlcNAc supplementation assay in fibroblasts. (a) Measurement of very long-chain fatty acids (VLCFAs) in the lysate of patient fibroblasts normalized to the mean values of the control cell lysate revealed partially increased levels. (b) Staining of lipids in control and patient fibroblasts with Oil Red O presented with an increase in pronounced coloration in the case of the patient's cells indicating an accumulation of neutral lipids. (c) Nanoelectrospray ionization tandem mass spectrometry of cellular lipids revealed significant alterations concerning ceramide (Cer), phosphatidylethanolamine plasmalogen (PE-P), and triacylglycerol (TAG) within the main lipid classes. Abbreviations: CE, cholesteryl ester; Chol, cholesterol; DAG, diacylglycerol; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PI, phosphatidylinositol; PS, phosphatidylserine; SM, sphingomyelin; Hex2Cer, dihexosylceramide; HexCer, hexosylceramide; LPC, lysophosphatidylcholine; PA, phosphatidic acid; PG, phosphatidylglycerol. (d) Reduced expression of ATP5H combined with a decreased ATP level (e) hint to a general mitochondrial impairment in SLC35A1-CDG. (f) Reduced proliferation due to decreased growth rate and increased doubling time in the patient's cells can be improved by 10 mM GlcNAc as a cell culture supplement.

Figure Legend Snippet: Metabolic measurements and GlcNAc supplementation assay in fibroblasts. (a) Measurement of very long-chain fatty acids (VLCFAs) in the lysate of patient fibroblasts normalized to the mean values of the control cell lysate revealed partially increased levels. (b) Staining of lipids in control and patient fibroblasts with Oil Red O presented with an increase in pronounced coloration in the case of the patient's cells indicating an accumulation of neutral lipids. (c) Nanoelectrospray ionization tandem mass spectrometry of cellular lipids revealed significant alterations concerning ceramide (Cer), phosphatidylethanolamine plasmalogen (PE-P), and triacylglycerol (TAG) within the main lipid classes. Abbreviations: CE, cholesteryl ester; Chol, cholesterol; DAG, diacylglycerol; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PI, phosphatidylinositol; PS, phosphatidylserine; SM, sphingomyelin; Hex2Cer, dihexosylceramide; HexCer, hexosylceramide; LPC, lysophosphatidylcholine; PA, phosphatidic acid; PG, phosphatidylglycerol. (d) Reduced expression of ATP5H combined with a decreased ATP level (e) hint to a general mitochondrial impairment in SLC35A1-CDG. (f) Reduced proliferation due to decreased growth rate and increased doubling time in the patient's cells can be improved by 10 mM GlcNAc as a cell culture supplement.

Techniques Used: Control, Staining, Mass Spectrometry, Expressing, Cell Culture

Incubation:

Article Title: A Novel Missense Variant in Ultrarare SLC35A1-CDG Alters Cellular Glycosylation, Lipid, and Energy Metabolism Without Affecting CDG Serum Markers
Article Snippet: .. Primary antibodies for detection of SLC35A1 (Proteintech, BIOZOL, MyBioSource), ADAMTS13, ATP5H, GP130 (all Thermo Fisher), GAPDH, histone 2B (H2B), H2B-GlcNAc (all Abcam), ICAM1 (Sigma), OGA, OGT, TGN46 (all Proteintech), O-GlcNAcylation (Santa Cruz), and HA-Tag (Sigma) were used in 1:1000 dilution, β -ACTIN (Sigma) in 1:10,000 dilution, and incubated overnight at 4°C under constant movement. .. Primary antibodies for detection of SLC35A1 (Proteintech, BIOZOL, MyBioSource), ADAMTS13, ATP5H, GP130 (all Thermo Fisher), GAPDH, histone 2B (H2B), H2B-GlcNAc (all Abcam), ICAM1 (Sigma), OGA, OGT, TGN46 (all Proteintech), O-GlcNAcylation (Santa Cruz), and HA-Tag (Sigma) were used in 1:1000 dilution, β -ACTIN (Sigma) in 1:10,000 dilution, and incubated overnight at 4°C under constant movement.

Western Blot:

Article Title: The SUMO-interacting motif in NS2 promotes adaptation of avian influenza virus to mammals
Article Snippet: Cell lysate (100 μl) was saved as an input control, and the remaining cell lysate was incubated with Glutathione MagBeads (GenScript, L00327) overnight at 4°C. .. The following antibodies were used: rabbit anti-GST (1:2000 for WB; GenScript, A00097), rabbit anti-Flag (1:2000 for WB; Sigma-Aldrich, F7425), rabbit anti-V5 (1:2000 for WB; Proteintech, 14440-1-AP), mouse anti-His (1:5000 for WB; Proteintech, 66005-1-Ig), rabbit anti-ACTB (β-Actin) (1:10,000 for WB; Abclonal, AC026), mouse anti-ACTB (1:2000 for WB; Abclonal, AC004), rabbit anti-Flag (1:5000 for WB; Sigma-Aldrich, F7425), rabbit anti-IAV PB2 (1:5000 for WB; GeneTex, GTX125926), mouse anti-human ANP32A(1:5000 for WB; Proteintech, 67687-1-Ig), rabbit anti-human ANP32B (1:5000 for WB; Proteintech, 10843-1-AP), mouse anti-IAV PA (1:5000 for WB; prepared in our laboratory), mouse anti-IAV PB2 (1:1000 for WB; prepared in our laboratory), and mouse anti-IAV NP (1:1000 for WB; prepared in our laboratory). ..

Recombinant:

Article Title: Protein arginine methyltransferase 5 mediates arginine symmetric dimethylation of influenza A virus PB2 and supports viral replication.
Article Snippet: All viruses were propagated using 10‐day‐old specific pathogen‐free (SPF) chicken embryos purchased from Beijing Boehringer Ingelheim Vital Biotechnology Co. Ltd. (Beijing, China) and then titrated by 50% tissue culture infectious dose (TCID50) analysis on MDCK cells. .. Antibodies and reagents used in this study: anti‐PRMT5 recombinant rabbit monoclonal antibody (mAb) (Cat No. ET1609‐43; HUABIO); Pan‐Symmetric Di‐Methyl Arginine Motif Rabbit mAb (Cat No. A20794; ABclonal); anti‐GAPDH mouse mAb (Cat No. 60004‐1‐Ig; Proteintech); anti‐IAV NP, PA, PB1, PB2, M1, M2, and HA rabbit polyclonal antibodies (pAb) (Cat No. GTX125989, GTX118991, GTX125923, GTX125926, GTX125928, GTX125951, and GTX127357; GeneTex); anti‐Flag mouse mAb (Cat No. F1804; Sigma); anti‐HA mouse mAb (Cat No. M180‐3, MBL); anti‐Lamin A/C rabbit pAb (Cat No. ET7110‐12; HUABIO); anti‐Clathrin heavy chain rabbit pAb (Cat No. A14323; ABclonal); anti‐Cy3 goat anti‐rabbit IgG (H+L) (Cat No. AS007; ABclonal); FITC goat anti‐mouse IgG (H+L) (Cat No. AS001; ABclonal); 4′, 6‐diamidino‐ 2‐phenylindole (DAPI) (Cat No. C1002; Beyotime); and Protein A/G Magnetic Beads (Cat No. HY‐K0202; MCE). ..