Analysis of respiratory mucus glycoproteins in asthma: a detailed study from a patient who died in status asthmaticus.
Airway mucus from asthmatics is often unusually solid. The death of a patient in status asthmaticus allowed the collection of 28 g of abnormal airway mucus at autopsy. Its chemical and physical properties were studied to reveal differences from more normal airway mucus. The gel plug taken from the airways could be dispersed in 6 M guanidinium chloride, but it took > 1 wk and 700 ml of extractant to disperse 3 g of exudate completely. In contrast, treatment with 10 mM dithiothreitol, which reduces disulfide bonds, dispersed the gel within seconds. Mucins accounted for 25% of the non-dialyzable material in the gel, while DNA constituted < 1% and proteoglycans could not be detected. The mucins were similar in architecture and general composition to other respiratory mucins and were present at a high concentration (approximately 40 mg/ml). The majority of mucins were of extreme size (mean M(r) 30-40 x 10(6)) and slow to dissolve, but sequential extraction experiments on the gel exudate demonstrated a proportion of mucins (15%), the most readily extracted, which had a higher density, 1.45-1.55 g/ml, a lower M(r) (11.5 x 10(6)) and were markedly more acidic than the bulk of the mucins. Both major and minor mucin populations were extremely heterogeneous in mass distribution. Electron microscopy of the major mucin species demonstrated extensive networks of molecules many microns in length. The major mucin species was distinctly less acidic than mucins previously described from either normal or diseased airways. Amino acid analysis of fractions across the charge distribution suggested the presence of at least two different mucin proteins occurring as distinct glycoforms.
Sheehan
J K
JK
Division of Biochemistry, School of Biological Sciences, Manchester University, United Kingdom.
Richardson
P S
PS
Fung
D C
DC
Howard
M
M
Thornton
D J
DJ
D005782:Gels; D006023:Glycoproteins / Q000032:analysis* / Q000302:isolation & purification / Q000648:ultrastructure; D006801:Humans; D008168:Lung / Q000737:chemistry*; D008297:Male; D008854:Microscopy, Electron; D008875:Middle Aged; D008970:Molecular Weight; D009077:Mucins / Q000032:analysis / Q000302:isolation & purification / Q000648:ultrastructure; D009093:Mucus / Q000737:chemistry*; D013183:Sputum / Q000737:chemistry; D013224:Status Asthmaticus / Q000503:physiopathology*
D005782:Gels; D006023:Glycoproteins; D009077:Mucins