Molecular analysis of the gene of the alpha 1-antitrypsin deficiency variant, Mnichinan

Abstract

Mnichinan, a variant of alpha 1-antitrypsin (alpha 1-AT) was detected in a Japanese individual with serum alpha 1-AT deficiency (18 mg/dl), associated with aggregated alpha 1-AT molecules in the hepatocytes. Cloning and sequencing of the 10,627-bp-long region containing the Mnichinan gene and the normal M1(Val213) alpha 1-AT gene revealed all five exons of the Mnichinan gene to be identical with the M1(Val213) alpha 1-AT gene, except for two changes: a TTC trinucleotide deletion in the codon for amino acid Phe52 and a G-A substitution, by which the normal Gly148 (GGG) became Arg148 (AGG). Dot blot analysis of the polymerase chain-reaction-amplified DNA derived from the proband and other family members showed both mutations to be associated with an alpha 1-AT deficiency phenotype. Ninety-eight alpha 1-AT alleles were all negative for both changes. Comparison of the region, except for five exons between the Mnichinan and M1(Val213) genes, demonstrated one base difference in the 5' flanking region and 14 base changes in the introns. All exon-intron junctions were identical, and base changes in the 5' flanking region did not seem significant. The G-A substitution in codon 148 of the Mnichinan gene could not be responsible for the alpha 1-AT deficiency phenotype because Arg- and not Gly- was located at the corresponding position of the protein C inhibitor belonging to the serine protease inhibitor superfamily. The deletion of Phe52 may cause the newly synthesized alpha 1-AT protein to aggregate, resulting in alpha 1-AT deficiency. Comparison of the alpha 1-AT gene sequences available indicated that the C-T substitution at the CpG dinucleotide has an important role in generation of variants and nucleotide changes in the noncoding regions of the alpha 1-AT gene.