caa a22 4500 465764258 CHVBK 20180323111909.0 cr unu---uuuuu 170327e19900801xx s 000 0 eng 10.1007/BF00914088 doi (NATIONALLICENCE)springer-10.1007/BF00914088 Neutral and acidic human tracheobronchial mucin [Elektronische Daten] Isolation and characterization of core protein [S. Bhattacharyya, B. Veit, B. Manna, J. Enriquez, M. Walker, A. Khorrami, B. Kaufman] Human bronchial mucin from a patient suffering from chronic bronchitis was solubilized in aqueous solution containing sodium azide and protease inhibitors and purified by Sepharose 4B and 2B column chromatography. The mucin was further purified by cesium bromide density gradient centrifugation. Sodium dodecyl sulfate-polyacrylamide gel (7.5%) electrophoresis of this material showed high-molecular-weight mucin component(s) at the top of the gel. Chemical analysis of this preparation indicated a typical mucin profile of amino acids and carbohydrates. Ion-exchange chromatography resulted in resolution of the purified mucin into neutral and acidic fractions. Comparison of the chemical composition of these two fractions showed higher mole percentage of threonine, serine, sialic acid, and sulfate in the acidic fraction. Chemical deglycosylation of the purified mucin preparation with trifluoromethane sulfonic acid was carried out at 20°C for 3 1/2 h. Sialic acid, fucose, galactose, and W-acetylglucosamine were completely removed, whereas traces ofN- acetylgalactosamine were still detected. High-pressure liquid chromatography of the deglycosylated products from native, neutral, and acidic mucin preparations resulted in a principal peptide, P1, with identical amino acid composition. Cyanogen bromide (CNBr) treatment of the peptide P1 from neutral and acidic mucins and subsequent fractionation of the fragments by high-pressure liquid chromatography resulted in similar peptide profiles. The P1 peptide fraction was further subjected to high-pressure liquid chromatography in a second solvent system, which resulted in two peaks, P1a and P1b. Gel filtration of both peptides in 6 M guanidine hydrochloride indicated a single peak with molecular weight of approximately 97 kDa. The amino acid profile of the two peptides was dominated by high levels of threonine, serine, and proline, which combined accounted for nearly 39% of the total residues, and in most respects, the profile resembled that of native mucin. End-group analysis of the peptide P1a indicated a blocked N-terminus, whereas serine was found to be the N-terminal amino acid in the peptide P1b. Rabbit antibodies prepared against the peptide P1 from native tracheal mucin reacted strongly with neutral and acidic mucin as well as the mucin from human colon. Both neutral and acidic human tracheal mucins were immunologically reactive with mouse monoclonal antibody HMPFG-2, which was prepared against human mammary mucin. However, the response of this antibody to human colonic mucin was rather weak. Plenum Publishing Corporation, 1990 Bhattacharyya S. Department of Clinical Investigation, William Beaumont Army Medical Center, El Paso, Texas aut Veit B. Department of Clinical Investigation, William Beaumont Army Medical Center, El Paso, Texas aut Manna B. Department of Clinical Investigation, William Beaumont Army Medical Center, El Paso, Texas aut Enriquez J. Department of Clinical Investigation, William Beaumont Army Medical Center, El Paso, Texas aut Walker M. National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina aut Khorrami A. Department of Biochemistry, Duke University Medical Center, Durham, North Carolina aut Kaufman B. Department of Biochemistry, Duke University Medical Center, Durham, North Carolina aut Inflammation Kluwer Academic Publishers-Plenum Publishers 14/4(1990-08-01), 355-373 0360-3997 14:4<355 1990 14 10753 https://doi.org/10.1007/BF00914088 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00914088 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Bhattacharyya S. Department of Clinical Investigation, William Beaumont Army Medical Center, El Paso, Texas aut NATIONALLICENCE 700 1- Veit B. Department of Clinical Investigation, William Beaumont Army Medical Center, El Paso, Texas aut NATIONALLICENCE 700 1- Manna B. Department of Clinical Investigation, William Beaumont Army Medical Center, El Paso, Texas aut NATIONALLICENCE 700 1- Enriquez J. Department of Clinical Investigation, William Beaumont Army Medical Center, El Paso, Texas aut NATIONALLICENCE 700 1- Walker M. National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina aut NATIONALLICENCE 700 1- Khorrami A. Department of Biochemistry, Duke University Medical Center, Durham, North Carolina aut NATIONALLICENCE 700 1- Kaufman B. Department of Biochemistry, Duke University Medical Center, Durham, North Carolina aut NATIONALLICENCE 773 0- Inflammation Kluwer Academic Publishers-Plenum Publishers 14/4(1990-08-01), 355-373 0360-3997 14:4<355 1990 14 10753 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer