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. 1973 Jul;70(7):2124–2128. doi: 10.1073/pnas.70.7.2124

Chemical Synthesis and Biochemical Properties of Peptide Fragments of Apolipoprotein-Alanine

James T Sparrow 1, Antonio M Gotto Jr 1, Joel D Morrisett 1
PMCID: PMC433680  PMID: 4352973

Abstract

Apolipoprotein-alanine is an apolipoprotein isolated from very-low-density lipoproteins of human plasma. This protein contains 79 amino acids and binds phosphatidyl choline. Four fragments of this molecule corresponding to sequence positions 41-79 (I), 48-79 (II), 55-79 (III), and 61-79 (IV) have been synthesized by standard solid-phase methods. The resulting peptides were cleaved from the resin and deblocked with liquid HF, then purified by chromatography on Sephadex G-50 and DEAE-cellulose. Each purified peptide eluted as a single, symmetrical peak, exhibited a single band on polyacrylamide gel electrophoresis, and gave an amino-acid analysis in good agreement with the theoretical value. Circular dichroism studies indicated that only fragments I and II became more helical in the presence of phosphatidyl choline and significantly inhibited the reactivation of delipidated β-hydroxybutyrate dehydrogenase (EC 1.1.1.30), an enzyme that requires phosphatidyl choline for activity. When subjected to ultracentrifugation at density 1.064 g/ml in the presence of phosphatidyl choline, fragments I, II, III, and IV floated to the top of the tube to the extent of 85, 50, 13, and 9%, respectively. These results indicate that residues 55-79 do not contain the minimum determinants required for the binding of phospholipid. However, extension of the peptide's N-terminus by seven residues produces a molecule that does bind phosphatidyl choline.

Keywords: solid-phase peptide synthesis, lipoproteins, phospholipid, circular dichroism, ultracentrifugation, β-hydroxybutyrate apodehydrogenase

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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