PMID: 222336Apr 25, 1979

A novel method for converting apolipoprotein B, the major protein moiety of human plasma low density lipoproteins, into a water-soluble protein

Biochimica Et Biophysica Acta
S S Huang, D M Lee

Abstract

We have developed a new technique to solubilize apolipoprotein B (ApoB) in aqueous solutions. The procedure involves stirring ApoB in 6 M guanidine/20 mM NH4Cl/NH4OH in the presence of cupric ammonia complexes at pH 9.7 for 20 h in N2, and then removing these reagents by a series of dialysis in N2. The resulting Cu(NH3)4(2)+-treated (Cu2+-treated) ApoB is soluble in aqueous buffers of pH above 8.3 or below 3. Parallel experiments carried out on control proteins, human albumin, alpha-lactalbumin, and insulin, indicated no change in molecular weight and no creation of a new NH2-terminal amino acid after Cu2+-treatment. By Edman degradation, the Cu2+-treated ApoB showed no detectable NH2-terminal amino acid. These results showed that the mechanism of Cu2+-solubilization of ApoB was not due to the cleavage of peptide bonds. Electrophoresis on urea-polyacrylamide gel, Cu2+-treated ApoB showed the same number of bands as the non-treated ApoB in the separating gels (7%) near the cathode, suggesting the heterogeneity of ApoB. In SDS-polyacrylamide gel (10%), the reduced and Cu2+-treated ApoB migrated with the similar mobilities to the monomer or dimer of human albumin. Antibodies raised against Cu2+-treated ApoB gave at least two immun...Continue Reading

References

Apr 1, 1978·Biochemical Medicine·C H Chen, F Aladjem
Jul 1, 1978·Proceedings of the National Academy of Sciences of the United States of America·J SchuhR H Haschemeyer
Apr 21, 1975·Biochemical and Biophysical Research Communications·P SepulvedaJ Tang
Sep 23, 1974·Biochemical and Biophysical Research Communications·C H Chen, F Aladjem
Aug 1, 1970·Proceedings of the National Academy of Sciences of the United States of America·J P KaneR J Havel
Jan 1, 1969·Proceedings of the National Academy of Sciences of the United States of America·A ScanuK Kothary
Sep 27, 1967·Biochemical and Biophysical Research Communications·B Shore, V Shore
Dec 15, 1996·Circulation·S SchäferB E Strauer

Citations

Jul 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·A J LusisM C Schotz
Oct 11, 1985·Nucleic Acids Research·M MehrabianE Mendiaz
Aug 1, 1980·Thrombosis Research·H MaedaH Uzawa
Jun 18, 1985·European Journal of Biochemistry·M WettestenS O Olofsson

Related Concepts

Apolipoproteins
Copper
SDS-PAGE
Immunodiffusion Measurement
Guanidines
LDL-1
Study, Methodological

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