An analysis of the interaction between mouse apolipoprotein B100 and apolipoprotein(a).
The assembly of lipoprotein(a) (Lp(a)) involves an initial noncovalent interaction between apolipoprotein (apo) B100 and apo(a), followed by the formation of a disulfide bond between apoB100 cysteine 4326 and apo(a) cysteine 4057. The structural features of apoB100 that are required for its noncovalent interaction with apo(a) have not been fully defined. To analyze that initial interaction, we tested whether apo(a) could bind noncovalently to two apoB proteins that lack cysteine 4326: mouse apoB100 and human apoB100-C4326G. Our experiments demonstrated that both mouse apoB and the human apoB100-C4326G bind noncovalently to apo(a). We next sought to gain insights into the apoB amino acid sequences required for the interaction between apoB100 and apo(a). Previous studies of truncated human apoB proteins indicated that the carboxyl terminus of human apoB100 (amino acids 4330-4397) is important for Lp(a) assembly. To determine whether the carboxyl terminus of mouse apoB100 can interact with apo(a), transgenic mice were produced with a mutant human apoB gene construct in which human apoB100 amino acids 4279-4536 were replaced with the corresponding mouse apoB100 sequences and tyrosine 4326 was changed to a cysteine. The mutant apoB100 bound to apo(a) and formed bona fide disulfide-linked Lp(a), but Lp(a) assembly was less efficient than with wild-type human apoB100. The fact that Lp(a) assembly was less efficient with the mouse apoB sequences provides additional support for the notion that sequences in the carboxyl terminus of apoB100 are important for Lp(a) assembly.
Cheesman
E J
EJ
Biochemistry Department, University of Otago, Dunedin 9001, New Zealand.
Sharp
R J
RJ
Zlot
C H
CH
Liu
C Y
CY
Taylor
S
S
Marcovina
S M
SM
Young
S G
SG
McCormick
S P
SP
D019943:Amino Acid Substitution; D000818:Animals; D053299:Apolipoprotein B-100; D001053:Apolipoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001055:Apolipoproteins B / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D053241:Apoprotein(a); D001665:Binding Sites; D003545:Cysteine; D006801:Humans; D007700:Kinetics; D017270:Lipoprotein(a) / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D055503:Protein Multimerization; D017384:Sequence Deletion
D053299:Apolipoprotein B-100; D001053:Apolipoproteins; D001055:Apolipoproteins B; D017270:Lipoprotein(a); D053241:Apoprotein(a); D003545:Cysteine