RESUMO
Since in a previous study we encountered a subject with an unusual split MG2 banding pattern, the aim of this study was to investigate the molecular basis of this observation. Submandibular/sublingual secretion was collected under resting and stimulated conditions and examined on Western blots probed with anti-MG2 antibodies or on gels stained with periodic acid-Schiff reagent. Genomic DNA was isolated and the N-, tandem repeat (TR), and C-terminal regions of MUC7 were amplified by PCR since MG2 is known to display a genetic polymorphism. Although the typical appearance of MG2 on blots and gels is a single 180 kDa band, salivary secretions from the subject exhibited doublet immunoreactive bands of approximately 180 and 125 kDa. Additionally, under resting conditions the 180 kDa band was predominant whereas upon stimulation the 125 kDa band became predominant. Genomic DNA analysis showed that MUC7 in the individual with split MG2 bands was not truncated and that the MUC7 genotype in this individual was (6/6) where both alleles encoded six TRs. The MG2 split banding pattern observed in this subject was not derived from proteolytic degradation of this salivary mucin in whole saliva or from genetic polymorphism. The expression of two isoforms of MG2 could in principle improve or reduce the activity of this key component of the oral host defense system.
Assuntos
Mucinas/genética , Mucinas/imunologia , Proteínas e Peptídeos Salivares/genética , Proteínas e Peptídeos Salivares/imunologia , Glândula Sublingual/imunologia , Glândula Submandibular/imunologia , Adulto , Western Blotting , Bandeamento Cromossômico , Feminino , Genótipo , Glicosilação , Humanos , Masculino , Peso Molecular , Polimorfismo Genético , Isoformas de Proteínas , Análise de Sequência de DNA , Glândula Sublingual/metabolismo , Glândula Submandibular/metabolismo , Sequências de Repetição em Tandem/genéticaRESUMO
Individuals with periodontitis exhibit differential expression of mucin-glycoprotein-2 (MG2), a protein encoded by the MUC7 gene. It is well known that MG2 exerts bactericidal activity as well as exhibiting genetic polymorphism involving a variable number of tandem repeats (VNTR). In the present study, we assessed the distribution of allelic variants of the MUC7 gene in 22 individuals with aggressive periodonitis, 68 with chronic periodonitis, and 87 without periodonitis. Oral mucosal cells were collected, the DNA was extracted, and specific primers were used to amplify the region encoding the MUC7 tandem repeats (TRs). Polymerase chain reaction products were subjected to electrophoresis and analyzed on polyacrylamide gels stained with silver nitrate. Although the percentage distribution of homozygosity (6-6TR) and heterozygosity (5-6TR) showed variation among the groups, the observed differences were not statistically significant (P > 0.05; Fisher's Exact Test). The present results indicate that the expression of different numbers of TRs in this salivary mucin in the oral environment does not interfere with the etiopathogenesis of aggressive or chronic periodontitis.
