title: Influence of an asparagine to lysine mutation at amino acid 3516 of apolipoprotein B on low-density lipoprotein receptor binding.
abstract: BACKGROUND
  Three mutations in the apolipoprotein B (apoB) gene have previously been established as important causes of impaired receptor binding of LDL and, hence, Familial Defective Apolipoprotein B 100 (FDB). Previously, undescribed mutations were sought.
  
  
  METHODS
  Using denaturing gradient gel electrophoresis for mutation detection, DNA from 1852 new patients was examined.
  
  
  RESULTS
  A previously undiscovered mutation was found in codon 3516, located between known FDB mutations at codons 3500 and 3531. The new mutation introduces a positively charged amino acid-lysine-while other FDB mutations remove a positively charged residue, arginine. The phenotype was intriguing, LDL derived from N3516K heterozygotes allowed only poor growth of an LDL cholesterol-dependent cell line. ApoB-100-specific antibody MB47 bound to LDL from N3516K heterozygotes with increased affinity indicating a probable conformational change caused by the substitution. In contrast to these results, a competitive displacement assay in fibroblasts showed normal (or better) binding affinity to LDL receptors and using dynamic laser scattering no preferential accumulation of 3516K LDL particles in plasma was found.
  
  
  CONCLUSIONS
  Discovery of the mutation and characterisation of N3516K LDL reveals another naturally occurring apoB mutation that influences conformation of LDL apoB and its interaction with the LDL receptor.
authors.affiliation: Department of Pathological Biochemistry, Glasgow Royal Infirmary, North Glasgow Hospital University NHS Trust, Fourth floor, Queen Elizabeth Building, Alexandra Parade, Glasgow G31 2ER, Scotland, UK. dairena.gaffney@climmed.gal.ac.uk
authors.forename: Dairena
authors.initials: D
authors.lastname: Gaffney
authors.forename: Clive R
authors.initials: CR
authors.lastname: Pullinger
authors.forename: Denis St J
authors.initials: DS
authors.lastname: O'Reilly
authors.forename: Michael S
authors.initials: MS
authors.lastname: Hoffs
authors.forename: Isobel
authors.initials: I
authors.lastname: Cameron
authors.forename: J Keith
authors.initials: JK
authors.lastname: Vass
authors.forename: Medha V
authors.initials: MV
authors.lastname: Kulkarni
authors.forename: John P
authors.initials: JP
authors.lastname: Kane
authors.forename: Verne N
authors.initials: VN
authors.lastname: Schumaker
authors.forename: Gerald F
authors.initials: GF
authors.lastname: Watts
authors.forename: Chris J
authors.initials: CJ
authors.lastname: Packard
mesh_terms: D000328:Adult; D001055:Apolipoproteins B / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D001216:Asparagine / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D008078:Cholesterol, LDL / Q000378:metabolism; D004252:DNA Mutational Analysis; D004797:Enzyme-Linked Immunosorbent Assay; D005091:Exons / Q000235:genetics; D005260:Female; D005787:Gene Frequency; D006579:Heterozygote; D006801:Humans; D007834:Lasers; D008239:Lysine / Q000235:genetics / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D020125:Mutation, Missense / Q000235:genetics*; D010375:Pedigree; D010641:Phenotype; D012150:Polymorphism, Restriction Fragment Length; D011485:Protein Binding; D011973:Receptors, LDL / Q000378:metabolism*
chemical_list: D001055:Apolipoproteins B; D008078:Cholesterol, LDL; D011973:Receptors, LDL; D001216:Asparagine; D008239:Lysine