Adhesive properties of carcinoembryonic antigen glycoforms expressed in glycosylation-deficient Chinese hamster ovary cell lines.

Carcinoembryonic antigen (CEA) is an oncofoetal cell surface glycoprotein that serves as an important tumour marker for colorectal and some other carcinomas. Its immunoglobulin-like structure places CEA within the immunoglobulin superfamily. CEA functions in several biological roles including homotypic and heterotypic (with other CEA family members) cell adhesion. Cell-cell interaction can be modulated by different factors, e.g., post-translational modifications such as glycosylation. The purpose of this study was to examine whether changes in carbohydrate composition of CEA oligosaccharides can influence homotypic (CEA-CEA) interactions. In order to modulate glycosylation of CEA we used two different glycosylation mutants of Chinese hamster ovary (CHO) cells, Lec2 and Lec8. Lec2 cells should produce CEA with nonsialylated N-glycans, while Lec8 cells should yield more truncated sugar structures than Lec2. Parental CHO (Pro5) cells and the glycosylation deficient mutants were stably transfected with CEA cDNA. All three CEA glycoforms, tested in a solid-phase cell adhesion assay, showed an ability to mediate CEA-dependent cell adhesion, and no qualitative differences in the adhesion between the glycoforms were observed. Thus, it may be assumed that carbohydrates do not play a role in homotypic adhesion, and the interactions between CEA molecules depend solely on the polypeptide structure

Single nucleotide polymorphisms in <i>A4GALT</i> spur extra products of the human Gb3/CD77 synthase and underlie the P1PK blood group system

<div><p>Contrary to the mainstream blood group systems, P1PK continues to puzzle and generate controversies over its molecular background. The P1PK system comprises three glycosphingolipid antigens: P^k, P1 and NOR, all synthesised by a glycosyltransferase called Gb3/CD77 synthase. The P^k antigen is present in most individuals, whereas P1 frequency is lesser and varies regionally, thus underlying two common phenotypes: P₁, if the P1 antigen is present, and P₂, when P1 is absent. Null and NOR phenotypes are extremely rare. To date, several single nucleotide polymorphisms (SNPs) have been proposed to predict the P₁/P₂ status, but it has not been clear how important they are in general and in relation to each other, nor has it been clear how synthesis of NOR affects the P₁ phenotype. Here, we quantitatively analysed the phenotypes and <i>A4GALT</i> transcription in relation to the previously proposed SNPs in a sample of 109 individuals, and addressed potential P1 antigen level confounders, most notably the red cell membrane cholesterol content. While all the SNPs were associated with the P₁/P₂ blood type and rs5751348 was the most reliable, we found large differences in P1 level within groups defined by their genotype and substantial intercohort overlaps, which shows that the P1PK blood group system still eludes full understanding.</p></div

Scatter plots of RBC anti-P1 and anti-NOR antibody binding capacities (A) and relative <i>A4GALT</i> transcript levels in individuals with different genotypes and 95% confidence intervals on the intercohort mean differences (B).

<p>Scatter plots of RBC anti-P1 and anti-NOR antibody binding capacities (A) and relative <i>A4GALT</i> transcript levels in individuals with different genotypes and 95% confidence intervals on the intercohort mean differences (B).</p

Statistics on the effect of rs5751348 SNP on the P1 antigen level.

<p>Statistics on the effect of rs5751348 SNP on the P1 antigen level.</p