N-terminal cleavage of the salivary MUC5B mucin. Analogy with the Van Willebrand propolypeptide?
Sequence similarities between the oligomeric mucins (MUC2, MUC5AC, MUC5B) and the von Willebrand factor suggest that they may be assembled in a similar way. After oligomerization, a fragment corresponding to the D1 and D2 domains is released from the von Willebrand factor. This cleavage does not appear to occur in pig submaxillary mucin, the only mammalian mucin in which this cleavage has been examined thus far, but whether other oligomeric mucins undergo N terminus proteolysis is not known. Antibodies recognizing the D1, D2, D3, and the first Cys domains in MUC5B were established and used to investigate to what extent proteolytic cleavage occurs within the N-terminal part of salivary MUC5B. The antibodies against the D1 and D2 domains identified a polypeptide corresponding in size to a MUC5B fragment generated by cleavage within the D' domain analogously with the von Willebrand factor propolypeptide. The antibodies did not recognize the main mucin population, suggesting that the major part of salivary MUC5B is subjected to this cleavage. An antibody recognizing the D3 domain was used to reveal a second cleavage site in the "soluble" but not in the "insoluble" MUC5B fraction: the first structural difference observed between soluble and insoluble salivary MUC5B. The identification of these cleavage events shows that the N-terminal sites for MUC5B oligomerization are present in the D3 domain and/or in domains located C-terminal to this part of the molecule.
Wickström
C
C
Mucosal Biology Group, Department of Cell and Molecular Biology, Lund University, S-22184 Lund, Sweden.
Carlstedt
I
I
D015153:Blotting, Western; D002498:Centrifugation; D002499:Centrifugation, Density Gradient; D002586:Cesium / Q000494:pharmacology; D002712:Chlorides / Q000494:pharmacology; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D019791:Guanidine / Q000494:pharmacology; D006801:Humans; D007150:Immunohistochemistry; D055257:Mucin-5B; D009077:Mucins / Q000737:chemistry* / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012463:Saliva / Q000378:metabolism*; D012469:Salivary Glands / Q000378:metabolism*; D014132:Trachea / Q000378:metabolism; D014841:von Willebrand Factor / Q000378:metabolism*
D002712:Chlorides; C524578:MUC5B protein, human; D055257:Mucin-5B; D009077:Mucins; D010455:Peptides; D014841:von Willebrand Factor; D002586:Cesium; C028019:cesium chloride; D019791:Guanidine