7 Circular dichroism (CD) parameters and secondary structur estimates of proteins

Yang, Jen Tsi; Wu, Chunxiang; Böhm, Gerald

doi:10.1007/10086139_60

Cited by 3 publications

(2 citation statements)

References 72 publications

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“…The far-UV spectra showed that both BSM and PGM lacked distinct and well-defined secondary structures, such as α-helices, β-sheets, or β-turns. , This is hardly surprising as mucins are mainly made up of the heavily glycosylated “Proline-Threonine-Serine” (PTS) repeat central domains, and the stiffness of glycans tend to hinder the formation of highly ordered structures. ,− Only the N- and C-terminal domains have been assigned to contain structural motifs determined from the peptide sequence, including cysteine knots, CysD domains and Von Willebrand factor binding domains that are involved in mucin–mucin interactions. ,,,, However, the contribution from them to CD spectra signals is not significant due to the dominance of the glycosylated central domain. The far-UV CD spectra of BSM showed a weak, yet distinct local positive maximum at ca.…”

Section: Resultsmentioning

confidence: 99%

Structural and Mechanical Properties of Thin Films of Bovine Submaxillary Mucin versus Porcine Gastric Mucin on a Hydrophobic Surface in Aqueous Solutions

et al. 2016

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The structural and mechanical properties of thin films generated from two types of mucins, namely bovine submaxillary mucin (BSM) and porcine gastric mucin (PGM) in aqueous environment were investigated with several bulk and surface analytical techniques. Both mucins generated hydrated films on hydrophobic polydimethylsiloxane (PDMS) surfaces from 2 spontaneous adsorption arising from their amphiphilic characteristic. But, BSM formed more elastic films than PGM at neutral pH condition. This structural difference was manifested from the initial film formation processes to the responses to shear stresses applied to the films.Acidification of environmental pH led to strengthening the elastic character of BSM films with increased adsorbed mass, whereas an opposite trend was observed for PGM films. We propose that this contrast originates from that negatively charged motifs are present for both the central and terminal regions of BSM molecule, whereas a similar magnitude of negative charges is localized at the termini of PGM molecule. Given that hydrophobic motifs acting as anchor are also localized in the terminal region, electrostatic repulsion between anchoring units of PGM molecules on nonpolar PDMS surface leads to weakening of the mechanical integrity of the films.

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Section: Resultsmentioning

confidence: 99%

Structural and Mechanical Properties of Thin Films of Bovine Submaxillary Mucin versus Porcine Gastric Mucin on a Hydrophobic Surface in Aqueous Solutions

et al. 2016

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“…BSM shows a far-UV CD spectrum that corresponds to a random coil structure as there are no detectable features of α-helices or β-sheets (Figure ). − …”

Section: Resultsmentioning

confidence: 99%

Proteolytic Degradation of Bovine Submaxillary Mucin (BSM) and Its Impact on Adsorption and Lubrication at a Hydrophobic Surface

et al. 2015

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The effects of proteolytic digestion on bovine submaxillary mucin (BSM) were investigated in terms of changes in size, secondary structure, surface adsorption, and lubricating properties. Two proteases with distinctly different cleavage specificities, namely trypsin and pepsin, were employed. SDS-PAGE analysis with staining for proteins and carbohydrate moieties showed that only the unglycosylated terminal regions of BSM were degraded by the proteases. Size exclusion chromatography (SEC) and dynamic light scattering (DLS) analyses indicated that tryptic digestion mainly led to the reduction in size, whereas pepsin digestion rather caused an increase in the size of BSM. Less complete cleavage in terminal peptide regions by pepsin and subsequent aggregation were possibly responsible for the increased size. Far-UV circular dichroism (CD) spectra of the protease-treated BSM showed a slight change in the secondary structure owing to the removal of terminal domains, but the overall random coil conformation adopted by the central glycosylated domain remained dominant and essentially unchanged. Surface adsorption properties as characterized by optical waveguide lightmode spectroscopy (OWLS) showed that tryptic digestion of BSM resulted in a decrease in the adsorbed mass, but pepsin digestion led to a slight increase in the adsorbed mass onto a hydrophobic surface compared to intact BSM. This is in agreement with the partial preservation of peptide segments in the terminal regions after pepsin digestion as confirmed by SEC and DLS studies. Despite a contrast in the adsorbed amount of the protease-treated BSMs onto the surface, both proteases substantially deteriorated the lubricating capabilities of BSM at a hydrophobic interface. The present study supports the notion that the terminal domains of BSM are critical to the adsorption and lubricating properties of BSM at hydrophobic interfaces.

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A Simplified Chromatographic Approach to Purify Commercially Available Bovine Submaxillary Mucins (BSM)

Madsen

Pakkanen

Duelund

et al. 2014

Preparative Biochemistry and Biotechnology

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In this study, a simple purification protocol is developed to reduce the bovine serum albumin (BSA) content in commercially available bovine submaxillary mucin (BSM). This involved purification of the BSM by one-column anion-exchange chromatography protocol resulting in BSM with greatly reduced BSA content and homogeneously distributed size, and in a high yield of ∼43% from BSM as received from the manufacturer. The purity and composition of commercially acquired BSM were assessed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometry, which verified that BSA is the most abundant nonmucinous protein component. The purification effect was evident from a significantly altered circular dichroism (CD) spectrum of BSM after anion-exchange chromatography.

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7 Circular dichroism (CD) parameters and secondary structur estimates of proteins

Cited by 3 publications

References 72 publications

Structural and Mechanical Properties of Thin Films of Bovine Submaxillary Mucin versus Porcine Gastric Mucin on a Hydrophobic Surface in Aqueous Solutions

Structural and Mechanical Properties of Thin Films of Bovine Submaxillary Mucin versus Porcine Gastric Mucin on a Hydrophobic Surface in Aqueous Solutions

Proteolytic Degradation of Bovine Submaxillary Mucin (BSM) and Its Impact on Adsorption and Lubrication at a Hydrophobic Surface

A Simplified Chromatographic Approach to Purify Commercially Available Bovine Submaxillary Mucins (BSM)

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