Mutation analysis of the human adipocyte-specific apM-I gene

Background The aim of this study was to analyse the human adipocyte-specifi c apM-1 gene for sequence variations. Methods Sequence analysis was performed in 344 randomly chosen blood sample s using a capillary sequencer. Results Whereas no mutations were detected in intronic regions and in 2.7 k b of the promoter, two sequence variations were found within the coding seq uence of apM-1. For both mutations, a polymerase chain reaction-(PCR) based restriction fragment length polymorphism (RFLP) analysis was developed, wh ich provided a rapid screening method. A conservative T --> G transition at nucleotide + 45 within exon-2 [Gly15Gly] was detected with an allelic freq uency of 0.9 for the wild-type allele and 0.1 for thr mutated allele. In ad dition, a missense point mutation at nucleotide + 331 within exon-3 [Tyr111 His] was detected with an allelic frequency of 0.97 for the wild-type allel e and 0.03 for thr mutated allele. This mutation replaces a tyrosine by an histidine within the carboxyterminal globular domain of apM-1. Concerning t he Gly15Gly polymorphism, the TT genotype was found in 275 subjects (79.9%) , the TG genotype in 67 subjects (19.5%) and the GG genotype in 2 subjects (0.6%): one with maturity onset diabetes of young age (MODY-diabetes) and o ne with Lipoatrophic Diabetes Syndrome (LPDS). Concerning the Tyr111His pol ymorphism, the TT genotype was found in 328 subjects (95.4%), the TC genoty pe in 15 subjects (4.3%) and the CC genotype in 1 subject (0.3%). Conclusion The existence of two yet unknown mutations within the apM-1 gene was demonstrated and RFLP analysis was established for rapid screening. We ll defined cohorts of patients are necessary to determine the putative role of apM-1 gene mutations in the pathogenesis of metabolic disorders.