Structural and binding characteristics of the carboxyl terminal fragment of apolipophorin III from Manduca sexta

Biochemistry
Vasanthy NarayanaswamiRobert O Ryan

Abstract

The molecular basis of the interaction of apolipophorin III (apoLp-III), an exchangeable apolipoprotein from hemolymph of the sphinx moth. Manduca sexta, with lipoprotein surfaces and phospholipids was studied by investigating the structural and binding properties of the C-terminal fragment of the native protein. A 4K peptide, corresponding to the terminal helical segment of the native protein, was generated by cyanogen bromide treatment, purified by gel filtration and reverse-phase HPLC, and characterized by N-terminal sequencing and amino acid and mass spectrometric analysis. Circular dichroism (CD) spectroscopy of the peptide in buffer indicated a predominantly unstructured state while addition of trifluoroethanol (TFE), a helix-inducing agent, resulted in an alpha-helical structure. Sedimentation equilibrium studies revealed that the 4K peptide was monomeric in buffer. The 4K peptide assumed an alpha-helical conformation in the presence of sodium dodecyl sulfate (SDS) and lysolecithin, but was unstructured in the presence of dimyristoylphosphatidylcholine, either when added to preformed vesicles or upon cosonication, indicating an ability to bind to detergent micelles but not to phospholipid bilayers. Unlike native apoLp-II...Continue Reading

Citations

Apr 1, 1998·Protein Science : a Publication of the Protein Society·J WangRobert O Ryan
Jul 7, 1999·Biochimica Et Biophysica Acta·E GoormaghtighJ M Ruysschaert
Dec 22, 1999·Biochimica Et Biophysica Acta·Vasanthy Narayanaswami, Robert O Ryan
Jul 11, 2006·Archives of Biochemistry and Biophysics·Leonardo J LeonPaul M M Weers
Dec 1, 1999·Developmental and Comparative Immunology·A E Halwani, G B Dunphy
Jun 19, 2017·Protein Expression and Purification·Daniel E SalleePaul M M Weers
Jan 1, 1996·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·Robert O Ryan

Related Concepts

Apolipophorin-II
Metazoa
Apolipoproteins
Circular Dichroism, Vibrational
LDL-1
Membranes, Artificial
Models, Structural
Plasma Protein Binding Capacity
Protein Conformation
Trifluoroethanol

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