Four different mutations in codon 28 of α spectrin are associated with structurally and functionally abnormal spectrin α1/74 in hereditary elliptocytosis

Hereditary elliptocytosis (HE) Sp α1/74 is a disorder associated with defective spectrin (Sp) heterodimer self-association and an abnormal tryptic cleavage of the 80-kD αI domain of Sp resulting in increased amounts of a 74-kD peptide. The molecular basis of this disorder is heterogeneous and mutations in codons 28, 46, 48, and 49 (codons 22, 40, 42, and 43 in the previous nomenclature which did not include the six NH2-terminal amino acids) have been reported. In this study we present data on seven unrelated HE Sp α1/74 kindred from diverse racial backgrounds in whom we identified four different mutations all occurring in exon 2 of α Sp at codon 28. Utilizing the polymerase chain reaction we established a CGT → CTT; Arg → Leu 28 mutation in one kindred of Arab/Druze origin. In two unrelated white kindred of English/European origin the substitution is CGT → AGT; Arg → Ser 28 and in two apparently unrelated white kindred from New Zealand, the mutation is CGT → TGT; Arg → Cys 28. Finally, in one American black kindred and in a black kindred from Ghana the mutation involves CGT → CAT; Arg → His 28. Allele specific oligonucleotide hybridization confirmed that the probands are heterozygous for the respective mutant alleles. All four point mutations abolished an Aha II restriction enzyme site which allowed verification of linkage of the mutation with HE Sp α1/74. Our results imply that codon 28 of α Sp is a "hot spot" for mutations and also indicate that Arg 28 is critical for the conformational stability and functional self association of Sp heterodimers.