Abstract
The binding property of Con A has been studied intensively and applied widely to glycoconjugates / glycobiology for over 80 years. However, its role and functional relationship of Con A with these mammalian structural units, glycotopes, N-glycan chains, as well as their polyvalent forms in N-glycoproteins involved in the Con A-glycan interactions have not been well defined and organized. In this study, the recognition factors involved in these interactions were analyzed by our well developed method- the enzyme linked lectinosorbent (ELLSA) and inhibition assay. Based on all the data obtained, it is concluded that Con A, as previously reported, has a relatively broad and wide recognition ability of the Manα1 → and Glcα1 → related glycans. It reacted not only strongly with yeast mannan and glycogens, but also bound well with a large number of mammalian N-glycans, including the N-glycans of rat sublingual gp (RSL), human Tamm-Horsfall glycoprotein (THGP), thyroglobulin and lactoferrin. The recognition specificity of Con A towards ligands, expressed by Molar Relative Potency (Molar R.P.), in a decreasing order is as follows: α1 → 3, α1 → 6 Mannopentaose (M5) and Biantennary N-linked core pentasaccharide (MDi) ≥ α1 → 3, α1 → 6 Mannotriose (M3) > Manα1 → 3Man (α1 → 3Mannobiose), Manα1 → 2Man (α1 → 2Mannobiose), Manα1 → 6Man (α1 → 6Mannobiose), Manα1 → 4Man (α1 → 4Mannobiose) > GlcNAcβ1 → 2Man (β1 → 2 N-Acetyl glucosamine-mannose) > Manα1 → /Glcα1 → > Man > Glc, while Gal / GalNAc were inactive. Furthermore, the Man related code system, in this study, is proposed to express by both numbers of Man and GlcNAcβ1 → branches (M3 to M9 / MMono to Penta etc.) and a table of three Manα1 → and Glcα1 → related biomasses of six recognition factors involved in the Con A-glycan interactions has also been demonstrated. These themes should be one of the most valuable advances since 1980s.
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Abbreviations
- C(G)BP:
-
Carbohydrate (Glycan) Binding Protein
- Con A:
-
Concanavalin A lectin
- Morniga-M:
-
Morus nigra lectin
- Lentil:
-
Lens culinaris lectin
- PSA:
-
Pea (Pisum sativum) lectin
- THGP:
-
Tamm-Horsfall glycoprotein
- HOC:
-
Cyst gps: glycoproteins isolated from human ovarian cyst fluid
- RSL:
-
Rat sublingual gp-major
- OSM:
-
Ovine submandibular gp-major
- PSM:
-
Porcine salivary mucin-major
- BSM:
-
Bovine submandibular gp-major
- Man:
-
D-manno pyranose
- Glc:
-
D-glucopyranose
- GlcNAc:
-
N-Acetylglucosamine
- Gal:
-
D-galactopyranose
- GalNAc:
-
N-Acetylgalactosamine
- GalN:
-
Galactosamine
- LFuc:
-
6-deoxy-L-galactopyranose
- Neu5Ac:
-
SA, NeuAc, sialic acid
- LacNAc:
-
N-Acetyllactosamine
- M 3 / M 5 :
-
The No. of subscript in Man indicates the No. of Man in the structures of the mammalian N-glycans
- M or M 3 :
-
Manα1 → 6(Manα1 → 3)Man, or α1 → 3, α1 → 6 Mannotriose
- M 5 :
-
Manα1 → 6(Manα1 → 3)Manα1 → 6(Manα1 → 3)Man, or α1 → 3, α1 → 6 Mannopentaose
- M Mono to Tetra :
-
The Mono to Tetra or more of subscript in Man indicates the No. of GlcNAcβ1 → or IIβ branches in the structures of the mammalian N-glycan chain
- Tn :
-
GalNAcα1 → Ser/Thr
- T :
-
Thomsen-Friedenreich disaccharide
- T α :
-
Galβ1 → 3GalNAcα1 →
- A :
-
GalNAcα1 → 3Gal
- A h :
-
GalNAcα1 → 3(LFucα1 → 2)Gal (Human blood group A glycotope)
- B :
-
Galα1 → 3Gal
- B h :
-
Galα1 → 3(LFucα1 → 2)Gal (Human blood group B glycotope)
- H :
-
LFucα1 → 2Gal (Human blood group O (H) glycotope)
- h :
-
Crypto LFucα1 → 2Gal
- I β :
-
Galβ1 → 3GlcNAcβ1 →, human blood group type I precursor sequence
- II β :
-
Galβ1 → 4GlcNAcβ, human blood group type II precursor sequence
- Le a :
-
Lewisa, Galβ1 → 3(Fucα1 → 4)GlcNAc
- Le b :
-
Lewisb, Fucα1 → 2Galβ1 → 3(Fucα1 → 4)GlcNAc
- Le x :
-
Lewisx, Galβ1 → 4(Fucα1 → 3)GlcNAc
- Le y :
-
Lewisy, Fucα1 → 2Galβ1 → 4(Fucα1 → 3)GlcNAc
- sLe a :
-
Sialyl Lewisa, NeuAcα2 → 3Galβ1 → 3(Fucα1 → 4)GlcNAc
- sLe x :
-
Sialyl Lewisx, NeuAcα2 → 3Galβ1 → 4(Fucα1 → 3)GlcNAc
- C :
-
GlcNAcβ1 → 4GlcNAc, chitin disaccharide
- C f :
-
GlcNAcβ1 → 4(LFucα1 → 6)GlcNAc
- RFs:
-
Six Recognition Factors have been selected and defined- RF-1 to RF-6 or RF-(i) to (vi)
- Molar R.P.:
-
Molar Relative Potency for the avidities and/or intensities of RF-1 to RF-5 or RF-(i) to RF-(v)
- Mass R.P.:
-
Mass Relative Potency for the avidity and/or intensity of RF-6 or RF-(vi)
- ELLSA:
-
Enzyme linked lectinosorbent assay
- ELLISA:
-
Inhibition assay
- TBS:
-
Tris-HCl buffered saline
- TBS-T:
-
TBS with Tween 20
- PBS:
-
Phosphate-buffered saline
- MMCO:
-
Molecular mass cut off
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Acknowledgements
This work was supported by CGU research grant, BMRP 008, BMRP 530, and MICCs Forever Fund (in process), Kwei-San, Tao-yuan, Taiwan. The author would like to thanks Dr. Ming-Sung Tsai and Ms. Chien-Ching Wu for their immunochemical analyses, and Ms. Ko’s typing assistance.
Funding
It has been indicated at the end of our manuscript as this work was supported by CGU research grant, BMRP 008, BMRP 530, and MICCs Forever Fund (in process), Kwei-San, Tao-yuan, Taiwan.
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Wu, A.M. Roles of the structural units, glycotopes / mammalian N-glycans for Con A-glycan interactions, their codes, and their recognition factors. Glycoconj J 40, 587–608 (2023). https://doi.org/10.1007/s10719-023-10129-4
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DOI: https://doi.org/10.1007/s10719-023-10129-4