Glycoproteomic and Phenotypic Elucidation of B4GALNT2 Expression Variants in the SID Histo-Blood Group System
(2022) In International Journal of Molecular Sciences 23(7).- Abstract
The Sda histo-blood group antigen (GalNAcβ1-4(NeuAcα2-3)Galβ-R) is implicated in various infections and constitutes a potential biomarker for colon cancer. Sd(a−) individuals (2–4% of Europeans) may produce anti-Sda, which can lead to incompatible blood transfusions, especially if donors with the high-expressing Sd(a++)/Cad phenotype are involved. We previously reported the association of B4GALNT2 mutations with Sd(a−), which established the SID blood-group system. The present study provides causal proof underpinning this correlation. Sd(a−) HEK293 cells were trans-fected with different B4GALNT2 constructs and evaluated by immunostaining and glycoproteomics. The predominant SIDnull candidate allele with... (More)
The Sda histo-blood group antigen (GalNAcβ1-4(NeuAcα2-3)Galβ-R) is implicated in various infections and constitutes a potential biomarker for colon cancer. Sd(a−) individuals (2–4% of Europeans) may produce anti-Sda, which can lead to incompatible blood transfusions, especially if donors with the high-expressing Sd(a++)/Cad phenotype are involved. We previously reported the association of B4GALNT2 mutations with Sd(a−), which established the SID blood-group system. The present study provides causal proof underpinning this correlation. Sd(a−) HEK293 cells were trans-fected with different B4GALNT2 constructs and evaluated by immunostaining and glycoproteomics. The predominant SIDnull candidate allele with rs7224888:T>C (p.Cys406Arg) abolished Sda synthesis, while this antigen was detectable as N-or O-glycans on glycoproteins following transfection of wildtype B4GALNT2. Surprisingly, two rare missense variants, rs148441237:A>G and rs61743617:C>T, found in a Sd(a−) compound heterozygote, gave results similar to wildtype. To elucidate on whether Sd(a++)/Cad also depends on B4GALNT2 alterations, this gene was sequenced in five individuals. No Cad-specific changes were identified, but a detailed erythroid Cad glycoprotein profile was obtained, especially for glycophorin-A (GLPA) O-glycosylation, equilibrative nucleoside transporter 1 (S29A1) O-glycosylation, and band 3 anion transport protein (B3AT) N-glycosylation. In conclusion, the p.Cys406Arg β4GalNAc-T2 variant causes Sda-deficiency in humans, while the enigmatic Cad phenotype remains unresolved, albeit further characterized.
(Less)
- author
- Stenfelt, Linn LU ; Nilsson, Jonas ; Hellberg, Åsa LU ; Liew, Yew Wah ; Morrison, Jenny ; Larson, Göran and Olsson, Martin L. LU
- organization
- publishing date
- 2022
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- carbohydrate biosynthesis, erythrocyte, flow cytometry, gene expression, glycobiology, glycopeptide, glycoprotein, glycosyltransferase, mass spectrometry (MS), SID blood group system
- in
- International Journal of Molecular Sciences
- volume
- 23
- issue
- 7
- article number
- 3936
- publisher
- MDPI AG
- external identifiers
-
- pmid:35409292
- scopus:85127387083
- ISSN
- 1661-6596
- DOI
- 10.3390/ijms23073936
- language
- English
- LU publication?
- yes
- id
- 02c2d704-75f9-4460-bbeb-b0a538a14d21
- date added to LUP
- 2022-05-20 14:15:27
- date last changed
- 2024-09-19 21:43:08
@article{02c2d704-75f9-4460-bbeb-b0a538a14d21, abstract = {{<p>The Sd<sup>a</sup> histo-blood group antigen (GalNAcβ1-4(NeuAcα2-3)Galβ-R) is implicated in various infections and constitutes a potential biomarker for colon cancer. Sd(a−) individuals (2–4% of Europeans) may produce anti-Sd<sup>a</sup>, which can lead to incompatible blood transfusions, especially if donors with the high-expressing Sd(a++)/Cad phenotype are involved. We previously reported the association of B4GALNT2 mutations with Sd(a−), which established the SID blood-group system. The present study provides causal proof underpinning this correlation. Sd(a−) HEK293 cells were trans-fected with different B4GALNT2 constructs and evaluated by immunostaining and glycoproteomics. The predominant SID<sup>null</sup> candidate allele with rs7224888:T>C (p.Cys406Arg) abolished Sd<sup>a</sup> synthesis, while this antigen was detectable as N-or O-glycans on glycoproteins following transfection of wildtype B4GALNT2. Surprisingly, two rare missense variants, rs148441237:A>G and rs61743617:C>T, found in a Sd(a−) compound heterozygote, gave results similar to wildtype. To elucidate on whether Sd(a++)/Cad also depends on B4GALNT2 alterations, this gene was sequenced in five individuals. No Cad-specific changes were identified, but a detailed erythroid Cad glycoprotein profile was obtained, especially for glycophorin-A (GLPA) O-glycosylation, equilibrative nucleoside transporter 1 (S29A1) O-glycosylation, and band 3 anion transport protein (B3AT) N-glycosylation. In conclusion, the p.Cys406Arg β4GalNAc-T2 variant causes Sd<sup>a</sup>-deficiency in humans, while the enigmatic Cad phenotype remains unresolved, albeit further characterized.</p>}}, author = {{Stenfelt, Linn and Nilsson, Jonas and Hellberg, Åsa and Liew, Yew Wah and Morrison, Jenny and Larson, Göran and Olsson, Martin L.}}, issn = {{1661-6596}}, keywords = {{carbohydrate biosynthesis; erythrocyte; flow cytometry; gene expression; glycobiology; glycopeptide; glycoprotein; glycosyltransferase; mass spectrometry (MS); SID blood group system}}, language = {{eng}}, number = {{7}}, publisher = {{MDPI AG}}, series = {{International Journal of Molecular Sciences}}, title = {{Glycoproteomic and Phenotypic Elucidation of B4GALNT2 Expression Variants in the SID Histo-Blood Group System}}, url = {{http://dx.doi.org/10.3390/ijms23073936}}, doi = {{10.3390/ijms23073936}}, volume = {{23}}, year = {{2022}}, }