Discovery and consequences of apolipoprotein-epsilon(3Groningen): a G-insertion in codon 95/96 that is predicted to cause a premature stop codon.
BACKGROUND
We found an unexplained, persistent discrepancy between the outcomes of two apolipoprotein-E (apo-E) genotyping methods for a patient with features of familial dysbetalipoproteinaemia (FD). Polymerase chain reaction - restriction fragment length polymorphism resulted in the apo-epsilon(2)/epsilon(2) genotype, whereas minisequencing indicated apo-epsilon(2)/epsilon(3). The discrepancy was predicted to derive from a novel mutation.


METHODS
Sequencing of patient DNA, set-up of a mutation analysis method and establishment of mutation occurrence in 19 family members of the proband and investigation of its association with serum lipid indices.


RESULTS
Sequencing demonstrated a G-insertion in codon 95 or 96 ((95)AAG-(96-)GAG-->(95)AAG-(96)GGA-G) of the apo-epsilon(3) allele. The mutation, designated apo-epsilon(3Groningen), was predicted to cause a frameshift, a premature stop codon at codon 146 (AAG-->TAA) and the expression of a truncated apo-E protein, if any. Four family members with the apo-epsilon(3Groningen) were identified. Two family members with apo-epsilon(3)/epsilon(3Groningen) had serum lipid indices within reference ranges but low-serum apo-E. Three subjects with apo-epsilon(2)/epsilon(3Groningen), proband included, had serum cholesterol, triglycerides and calculated low-density lipoprotein-cholesterol levels above the reference ranges. Their electrophoresis pattern showed the classical broad-beta band, indicative of FD.


CONCLUSIONS
Apo-epsilon(3Groningen) heterozygosity is unlikely to precipitate FD, unless provoked by compound apo-epsilon(2) heterozygosity or other FD precipitating factors.
Dijck-Brouwer
D A Janneke
DA
Pathology and Laboratory Medicine, Groningen University Hospital, The Netherlands. d.a.j.brouwer@lc.umcg.nl
van Doormaal
Jasper J
JJ
Kema
Ido P
IP
Brugman
Astrid M
AM
Kingma
Anneke W
AW
Muskiet
Frits A J
FA
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001057:Apolipoproteins E / Q000235:genetics*; D018389:Codon, Nonsense / Q000235:genetics*; D004252:DNA Mutational Analysis; D005260:Female; D016368:Frameshift Mutation; D005838:Genotype; D006801:Humans; D006952:Hyperlipoproteinemia Type III / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D010375:Pedigree; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length
D001057:Apolipoproteins E; D018389:Codon, Nonsense