Application of lectin microarrays for the analysis of seminal plasma glycome.

We report application of lectin microarrays, exploiting simultaneous interaction of seminal plasma samples with multiple lectins of different sugar specificities to compare the glycomes of fertile and infertile men. The results indicate reduced lectin reactivity associated with decreased fertility, especially affecting oligozoospermic subjects and probably O-glycosylation. Lectin microarrays may become a potent tool for semen analysis in search of the association of glycosylation and male fertility.

Association of IgA secretory component sialylation with leucocytospermia of infertile men - a pilot study.

The aim of our pilot study was to check whether the differences in IgA secretory component (SC) sialylation are associated with leucocytospermia. In normozoospermic and leucocytospermic seminal plasmas, 78-kDa and 63-kDa SC immunoreactive bands were observed. The SC sialylation was analysed by lectin blotting, using sialo-specific lectins MAA (Maackia amurensis agglutinin) and SNA (Sambucus nigra agglutinin). Specific reactivity of 63-kDa SC with MAA and SNA was higher than 78-kDa SC in both analysed seminal groups. The analysis of seminal SC sialylation might be a valuable diagnosis tools for the evaluation of fertility problems related with leucocytospermia.

Fucosylation in synovial fluid as a novel clinical marker for differentiating joint diseases--a preliminary study.

OBJECTIVE: To investigate fucosylation of synovial fluid glycoproteins in patients with rheumatoid arthritis (RA), juvenile arthritis (JIA), gonarthrosis (GA) and reactive arthritis (ReA), referred to traumatized knee (TK). METHODS: Synovial fluid glycoproteins were separated by SDS-PAGE and either silver stained or blotted onto nitrocellulose and probed with the fucose-specific Aleuria aurantia lectin. Five bands were chosen for densitometric analysis. Total fucose content and density of fucosylated epitopes were analyzed. RESULTS: Fucose content was elevated in all patient groups and almost all bands, comparing to TK. The density of fucosylated epitopes was increased in the 42-kDa band of RA and JIA cases, and lowered in the 26-kDa band of RA and JIA, but not in GA. In all RA cases FR 42-kDa > FR 26-kDa. The relation was opposite in 8 out of 9 GA cases. CONCLUSION: The density of fucosylated epitopes differs significantly in particular glycoproteins of synovial fluid in joint diseases and may be of potential diagnostic value in differentiating diseases of inflammatory and degenerative origin.

The N-acetylgalactosamine and lactosamine specific lectin from Iris hybrida leaves.

The lectin isolated from the leaves of Iris hybrida binds specifically N-acetyl-galactosamine and lactose. Its molecule consists of two identical subunits bound by disulfide bonds. The lectin is a glycoprotein containing about 12% of sugars. It binds asialoglycoproteins containing complex type sugar chains. The binding is reduced by half at the concentration of 0.15 to 0.40 mM of the galactose containing disaccharides irrespectively to a type of galactose isomer. This indicates rather broad specificity of I. hybrida leaf lectin.

Carbohydrate structures of haptoglobin in sera of healthy people and a patient with congenital disorder of glycosylation.

Haptoglobin is one of acute phase glycoproteins often used as markers in glycopathology studies. In this work the oligosaccharide structures of haptoglobin from 'healthy' subjects have been studied in detail, taking into consideration the possible dependence of glycosylation on the phenotype. About 75% of charged haptoglobin glycans were of biantennary complex structure, and some of them lacked one terminal sialic acid molecule. Triantennary structures made up almost 25% of the charged glycans pool, and highly branched tetrasialylated oligosaccharides did not exceed 1%. The main difference between haptoglobin derived from the sample of pooled 44 sera and from the 2-2 phenotype individual concerned the relative content of trisialylated oligosaccharide with one 2-3 linked sialic acid residue. The oligosaccharide profile of haptoglobin derived from serum of a patient suffering from congenital disorder of glycosylation was compared to 'healthy' controls. It was shown, that four main glycans are identical in patient and 'normal' haptoglobins. Some alterations were found in the relative content of mono-, bi-, and trisialylated glycans as well as in the appearance of some tracely abundant oligosaccharides in haptoglobin of the patient with congenital disorder of glycosylation.

Haptoglobin glycoforms in a case of carbohydrate-deficient glycoprotein syndrome.

Alterations in haptoglobin (Hp) glycosylation were examined in the plasma of the first patient with carbohydrate-deficient glycoprotein syndrome (CDGS) who was described in Poland. Hp concentration in the CDGS patient plasma was low (240 mg/l) and the Hp phenotype was shown to be 2-2. Three glycoforms of the Hp beta subunit were observed in SDS-PAGE in CDGS. The densitometric analysis and molecular weight determinations suggested that 50% of glycoforms were fully glycosylated; 30% contained three out of four and 20% only two out of four glycan units compared to those that are present in Hp derived from healthy people. Results with lectins (concanavalin A and Sambucus nigra, Maackia amurensis and Alleuria aurantia agglutinins) indicate that all three glycoforms of beta subunit of CDGS-Hp contained biantennary complex glycans terminated with alpha2,6 bound sialic acid, but without fucose or alpha2,3 linked sialic acid. Hp glycosylation abnormalities described in this work suggest that this case was a type I carbohydrate-deficient glycoprotein syndrome.