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More than one disease process in chronic sinusitis based on mucin fragmentation patterns and amino Acid analysis.

Ali Mel-S, Pearson JP - Int J Otolaryngol (2015)

Bottom Line: Amino acid analysis was characteristic of mucins with high serine, threonine, and proline contents and reduction and proteolysis increased relative proportions of these amino acids.Conclusion.These data suggest that there may be different pathological processes occurring at the cellular level on chronic sinusitis.

View Article: PubMed Central - PubMed

Affiliation: Department of Otolaryngology, Mansoura University Hospital, Mansoura University, Mansoura 35516, Egypt ; Institute for Cell and Molecular Biosciences, Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne NE2 4HH, UK.

ABSTRACT
Objective. To characterise fragmentation patterns and amino acid composition of MUC2 and MUC5AC in chronic sinusitis. Methods. Antigenic identity of purified sinus mucins was determined by ELISA. Fragmentation patterns of a MUC5AC rich sample mucin were analysed by Sepharose CL-2B gel chromatography. Samples, divided into one MUC2 rich and one MUC5AC rich group, were subjected to sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and their amino acid contents were analysed. Results. Reduction, trypsin digestion, and papain digestion produced progressively smaller mucin species. On SDS-PAGE, digested MUC5AC rich mucin produced four distinct products. Amino acid analysis was characteristic of mucins with high serine, threonine, and proline contents and reduction and proteolysis increased relative proportions of these amino acids. MUC5AC rich mucins contained more protein than MUC2 rich mucins. Conclusion. Sinus mucin fragmentation produced mucin subunits and glycopeptide units of smaller molecular sizes which are likely to have lower viscoelastic properties. Applying this in vivo could alter mucus physical properties and biologic functions. Amino acid contents of MUC2 and MUC5AC mucins are different. This could be contributing to biological properties and functions of sinus mucins. These data suggest that there may be different pathological processes occurring at the cellular level on chronic sinusitis.

No MeSH data available.


Related in: MedlinePlus

Chromatographic profile of the purified fractionated sinus mucin sample (S1). Sepharose CL-2B gel column (125 × 2.5 cm) was eluted by upward flow with 0.2 M sodium chloride containing 0.02% (w/v) sodium azide and flow rate 18 mL/h. Calibration was firstly performed using 1% (w/v) dextran blue solution containing 0.05% (w/v) methyl orange. The glycoprotein content was estimated by the PAS solution assay. Vo and Vt are the void and total volumes, respectively.
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fig1: Chromatographic profile of the purified fractionated sinus mucin sample (S1). Sepharose CL-2B gel column (125 × 2.5 cm) was eluted by upward flow with 0.2 M sodium chloride containing 0.02% (w/v) sodium azide and flow rate 18 mL/h. Calibration was firstly performed using 1% (w/v) dextran blue solution containing 0.05% (w/v) methyl orange. The glycoprotein content was estimated by the PAS solution assay. Vo and Vt are the void and total volumes, respectively.

Mentions: Gel filtration was performed on MUC5AC rich sinus mucin sample (S1). The elution profiles are shown in Figure 1. Seventy-five percent of polymeric sinus mucin was excluded (Kav 0) and the remaining 25% spread into the partially included volume as a trailing edge. On reduction, sinus mucin eluted as two peaks: the first, representing 20% of the mucin, was excluded and the other 80% eluted as a partially included broad second peak (Kav 0.28). Trypsin digested mucin eluted as two included peaks. First peak made up 80% of the mucin (Kav 0.47) and the second (20%) constituted a shoulder of smaller size material (Kav 0.69). Papain digested polymeric mucin showed two included, partially separated peaks with Kav 0.57 and 0.78 accounting for 74% and 26% of loaded mucin, respectively.


More than one disease process in chronic sinusitis based on mucin fragmentation patterns and amino Acid analysis.

Ali Mel-S, Pearson JP - Int J Otolaryngol (2015)

Chromatographic profile of the purified fractionated sinus mucin sample (S1). Sepharose CL-2B gel column (125 × 2.5 cm) was eluted by upward flow with 0.2 M sodium chloride containing 0.02% (w/v) sodium azide and flow rate 18 mL/h. Calibration was firstly performed using 1% (w/v) dextran blue solution containing 0.05% (w/v) methyl orange. The glycoprotein content was estimated by the PAS solution assay. Vo and Vt are the void and total volumes, respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4321678&req=5

fig1: Chromatographic profile of the purified fractionated sinus mucin sample (S1). Sepharose CL-2B gel column (125 × 2.5 cm) was eluted by upward flow with 0.2 M sodium chloride containing 0.02% (w/v) sodium azide and flow rate 18 mL/h. Calibration was firstly performed using 1% (w/v) dextran blue solution containing 0.05% (w/v) methyl orange. The glycoprotein content was estimated by the PAS solution assay. Vo and Vt are the void and total volumes, respectively.
Mentions: Gel filtration was performed on MUC5AC rich sinus mucin sample (S1). The elution profiles are shown in Figure 1. Seventy-five percent of polymeric sinus mucin was excluded (Kav 0) and the remaining 25% spread into the partially included volume as a trailing edge. On reduction, sinus mucin eluted as two peaks: the first, representing 20% of the mucin, was excluded and the other 80% eluted as a partially included broad second peak (Kav 0.28). Trypsin digested mucin eluted as two included peaks. First peak made up 80% of the mucin (Kav 0.47) and the second (20%) constituted a shoulder of smaller size material (Kav 0.69). Papain digested polymeric mucin showed two included, partially separated peaks with Kav 0.57 and 0.78 accounting for 74% and 26% of loaded mucin, respectively.

Bottom Line: Amino acid analysis was characteristic of mucins with high serine, threonine, and proline contents and reduction and proteolysis increased relative proportions of these amino acids.Conclusion.These data suggest that there may be different pathological processes occurring at the cellular level on chronic sinusitis.

View Article: PubMed Central - PubMed

Affiliation: Department of Otolaryngology, Mansoura University Hospital, Mansoura University, Mansoura 35516, Egypt ; Institute for Cell and Molecular Biosciences, Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne NE2 4HH, UK.

ABSTRACT
Objective. To characterise fragmentation patterns and amino acid composition of MUC2 and MUC5AC in chronic sinusitis. Methods. Antigenic identity of purified sinus mucins was determined by ELISA. Fragmentation patterns of a MUC5AC rich sample mucin were analysed by Sepharose CL-2B gel chromatography. Samples, divided into one MUC2 rich and one MUC5AC rich group, were subjected to sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and their amino acid contents were analysed. Results. Reduction, trypsin digestion, and papain digestion produced progressively smaller mucin species. On SDS-PAGE, digested MUC5AC rich mucin produced four distinct products. Amino acid analysis was characteristic of mucins with high serine, threonine, and proline contents and reduction and proteolysis increased relative proportions of these amino acids. MUC5AC rich mucins contained more protein than MUC2 rich mucins. Conclusion. Sinus mucin fragmentation produced mucin subunits and glycopeptide units of smaller molecular sizes which are likely to have lower viscoelastic properties. Applying this in vivo could alter mucus physical properties and biologic functions. Amino acid contents of MUC2 and MUC5AC mucins are different. This could be contributing to biological properties and functions of sinus mucins. These data suggest that there may be different pathological processes occurring at the cellular level on chronic sinusitis.

No MeSH data available.


Related in: MedlinePlus