The Ile128Thr polymorphism influences stability and ligand binding properties of the microsomal triglyceride transfer protein.
The microsomal triglyceride transfer protein (MTTP) is essential for the assembly of VLDLs. We recently observed that a polymorphism in the MTTP promoter (-493G>T), which is in allelic association with an isoleucine-to-theronine substitution at position 128 (Ile128Thr) in the expressed protein, confers an increased risk of coronary heart disease. Two variant proteins comprising amino acids 16-297 of intact MTTP, MTTP(N)-Ile128 and MTTP(N)-Thr128, had similar native secondary structure content, as judged by circular dichroism. However, the thermal stability of MTTP(N)-Thr128 was greatly reduced, and this protein was also more extensively cleaved in limited proteolysis experiments compared with MTTP(N)-Ile128; both of these findings support a less compact fold. On adding LDL, which includes natively folded apolipoprotein B (apoB), decreased stability of the MTTP(N)-Thr128-LDL complex was observed compared with that of the MTTP(N)-Ile128-LDL complex. In a refined model of the N-terminal domain of MTTP, residue 128 is located in a surface-exposed position, in the same region as an identified MTTP binding site in the homologous apoB protein. Thus, the Ile128Thr polymorphism confers reduced structural stability, leading to decreased binding of MTTP to LDL particles. Because the major MTTP binding target on LDL is apoB, the Ile128Thr polymorphism could target the MTTP-apoB interaction.
Ledmyr
H
H
Atherosclerosis Research Unit, King Gustaf V Research Institute, Karolinska University Hospital, Stockholm, Sweden.
Ottosson
L
L
Sunnerhagen
M
M
Ehrenborg
E
E
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001055:Apolipoproteins B / Q000378:metabolism; D001483:Base Sequence; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D002918:Chymotrypsin; D002942:Circular Dichroism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004355:Drug Stability; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008024:Ligands; D008077:Lipoproteins, LDL / Q000378:metabolism; D008861:Microsomes / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011110:Polymorphism, Genetic; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D001055:Apolipoproteins B; D002352:Carrier Proteins; D018076:DNA, Complementary; D008024:Ligands; D008077:Lipoproteins, LDL; D011994:Recombinant Proteins; C106092:microsomal triglyceride transfer protein; D002918:Chymotrypsin