A mutation and an antibody that affect chemical cross-linking of low-density lipoprotein receptors on human fibroblasts.
Treatment of normal fibroblasts with the bifunctional cross-linking reagent DTSSP [3,3'-dithiobis(sulphosuccinimidylpropionate)] at 4 degrees C converted approximately 40% of the cell-surface low-density lipoprotein (LDL) receptors into a high-M(r) form, thought to represent receptor dimers. Preincubation of the cells with anti-(LDL receptor) monoclonal antibody 10A2 increased the proportion of surface receptors in the high-M(r) form after treatment with DTSSP at 4 degrees C to over 70%. Preincubation with LDL did not affect the proportion cross-linked, but prevented the increase produced by antibody 10A2. Cross-linking at 37 degrees C was less efficient than at 4 degrees C and was not affected by preincubation with antibody 10A2. Surface LDL receptors on fibroblasts from the homozygous familial hypercholesterolaemic subject MM were not cross-linked by DTSSP, confirming that the mutation had produced a change in the conformation of the receptor molecule. Taken together, the results suggest that normal LDL receptors on at least one region of the surface membrane may be loosely associated in some form of multimeric array which alters its alignment differently in response to antibody 10A2 and to cooling. Mutations that alter the tertiary structure of the receptors could affect LDL binding by disturbing the arrangement of the array.
Patel
D D
DD
MRC Lipoprotein Team, Hammersmith Hospital, London, U.K.
Soutar
A K
AK
Knight
B L
BL
D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D002478:Cells, Cultured; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D006937:Hypercholesterolemia / Q000235:genetics* / Q000378:metabolism*; D007700:Kinetics; D008077:Lipoproteins, LDL / Q000378:metabolism*; D009154:Mutation; D011973:Receptors, LDL / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D012867:Skin / Q000378:metabolism*; D013388:Succinimides / Q000494:pharmacology*
D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D008077:Lipoproteins, LDL; D011973:Receptors, LDL; D013388:Succinimides; C011240:dithiobis(succinimidylpropionate)