Respiratory mucins: identification of core proteins and glycoforms.
At least eight mucin apoproteins are expressed by the tracheobronchial epithelium, but it is not known which, if any, of these are major constituents of the respiratory secretions responsible for the formation of the mucus gel. To address this we have isolated mucins from normal, asthmatic and chronic bronchitic secretions. The asthmatic mucin reduced subunits were fractionated into four populations (I-IV) by anion-exchange HPLC. Amino acid and monosaccharide compositional analysis, as well as M(r) and size measurements, indicate that two of these populations (I and II) are glycoforms of the same or related apoprotein(s) and that the other populations contain two different apoproteins. A panel of antibodies and antisera recognizing the variable number tandem repeat (VNTR) of specific mucin apoproteins did not, as predicted, react with the glycosylated molecules, but after deglycosylation the majority of these probes (with the exception of those to MUC2, which were negative) reacted at a low level with each of the subunit populations. In contrast, an antiserum against a non-VNTR sequence of MUC5AC identified one of the populations (III) as the MUC5AC mucin. The MUC5AC reduced subunit had an M(r) of 2.2 x 10(6) and an RG (radius of gyration) of 57 nm. The genetic identities of the major mucin (populations I and II) and a minor component (population IV) were not established. The MUC5AC mucin was also identified as a major component in the pooled normal secretions from 20 individuals, whereas in a chronic bronchitic sample it was only a minor constituent. Furthermore, in all these different respiratory secretions the MUC5AC mucin appears as a similar biochemical entity, as assessed by Mono Q chromatography and agarose electrophoresis, suggesting that it may have a well-defined pattern of glycosylation in the respiratory tract.
Thornton
D J
DJ
Wellcome Trust Centre for Cell-Matrix Research, University of Manchester, School of Biological Sciences, U.K.
Carlstedt
I
I
Howard
M
M
Devine
P L
PL
Price
M R
MR
Sheehan
J K
JK
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal; D001249:Asthma / Q000503:physiopathology*; D001980:Bronchi / Q000378:metabolism; D001991:Bronchitis / Q000503:physiopathology*; D002241:Carbohydrates / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004848:Epithelium / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008168:Lung / Q000737:chemistry* / Q000378:metabolism; D046911:Macromolecular Substances; D051379:Mice; D008969:Molecular Sequence Data; D009077:Mucins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D012016:Reference Values; D013183:Sputum / Q000737:chemistry*; D014132:Trachea / Q000378:metabolism
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D046911:Macromolecular Substances; D009077:Mucins