title: Conformation of bovine submaxillary mucin layers on hydrophobic surface as studied by biomolecular probes.
abstract: In the present study, the conformational changes of bovine submaxillary mucin (BSM) adsorbed on a hydrophobic surface (polystyrene (PS)) as a function of concentration in bulk solution (up to 2mg/mL) have been investigated with biomolecular probe-based approaches, including bicinchoninic acid (BCA), enzyme-linked immunosorbent assay (EIA), and enzyme-linked lectin assay (ELLA). The conformation and hydrodynamic diameter of highly purified BSM molecules, as characterized by circular dichroism (CD) spectroscopy and dynamic light scattering (DLS), respectively, showed a slight, yet gradual coiling and compaction in response to the increase in BSM concentration in bulk solution. Adsorbed masses of BSM onto hydrophobic surface, as probe by BCA, showed a continuously increasing trend up to 2mg/mL. But, the signals from EIA and ELLA, which probe the concentration of available unglycosylated C-terminals and the central glycosylated regions, respectively, showed complicated non-linear responses with increasing surface concentration. The results from this study support the conventional amphiphilic, triblock model of BSM in the adsorption onto hydrophobic surface from aqueous solution. The biomolecular probe-based approaches employed in this study, however, provided further details on the conformational changes of BSM on surface, in particular the accessibility of glycosylated and unglycosylated domains with increasing surface concentration.
authors.affiliation: Department of Mechanical Engineering, Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark.
authors.forename: Kirsi I
authors.initials: KI
authors.lastname: Pakkanen
authors.affiliation: Department of Mechanical Engineering, Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark.
authors.forename: Jan B
authors.initials: JB
authors.lastname: Madsen
authors.affiliation: Department of Mechanical Engineering, Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark. Electronic address: seele@mek.dtu.dk.
authors.forename: Seunghwan
authors.initials: S
authors.lastname: Lee
mesh_terms: D000818:Animals; D002417:Cattle; D006031:Glycosylation; D057927:Hydrophobic and Hydrophilic Interactions; D009077:Mucins / Q000737:chemistry*; D011487:Protein Conformation; D054730:Protein Interaction Domains and Motifs; D012996:Solutions; D013363:Submandibular Gland / Q000378:metabolism
chemical_list: D009077:Mucins; D012996:Solutions
keywords: BSM; EIA; ELLA