Fluorescence studies of exchangeable apolipoprotein-lipid interactions. Superficial association of apolipophorin III with lipoprotein surfaces.

The Journal of Biological Chemistry
D SahooR O Ryan

Abstract

Apolipophorin III (apoLp-III) from the Sphinx moth, Manduca sexta, is an 18-kDa exchangeable apolipoprotein that reversibly associates with lipoprotein particles. In the absence of lipid, apoLp-III exists as an elongated bundle of five amphipathic alpha-helices. Upon lipid association, the protein is postulated to undergo a major conformational change, wherein the bundle opens around hinge loop regions, resulting in exposure of its hydrophobic interior. Fluorescence quenching techniques have been employed to study apoLp-III helix topography and spatial arrangement in phospholipid disc complexes and intact lipoprotein particles. Intrinsic fluorescence of the single tyrosine in apoLp-III was exploited to monitor the location of helix 5 in model disc complexes. To investigate other regions of the protein, site-directed mutagenesis was performed to introduce cysteine residues, replacing Asn-40 (helix 2, N40C) or Leu-90 (helix 3, L90C), thereby providing two mutant apoLp-IIIs, each with a single site for covalent attachment of the extrinsic fluorescent probe, N-(1-pyrene) maleimide. In the lipid-free state, pyrene-N40C- and pyrene-L90C-apoLp-III were highly accessible to the negatively charged aqueous quencher KI, yielding Ksv value...Continue Reading

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Citations

Jul 30, 2002·Journal of Molecular Biology·D SahooVasanthy Narayanaswami
Jul 7, 1999·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·C A FisherR O Ryan
Dec 22, 1999·Biochimica Et Biophysica Acta·Vasanthy Narayanaswami, R O Ryan
May 20, 2009·Biochimica Et Biophysica Acta·Lesley J VasquezPaul M M Weers
Feb 15, 2005·The Journal of Biological Chemistry·Jessica Drury, Vasanthy Narayanaswami
Dec 14, 2004·The Biochemical Journal·V RaussensVasanthy Narayanaswami
Jan 29, 2000·European Journal of Biochemistry·Vasanthy NarayanaswamiR O Ryan
Mar 28, 2003·The Journal of Biological Chemistry·Armelle Varnier AgasøsterAurora Martínez

Related Concepts

Apolipophorin-II
Alkylation
Metazoa
Apolipoproteins
Carrier Proteins
Lipoproteins
LDL-1
Turbidimetry
Type C Phospholipases
Protein Conformation

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