Intrinsic fluorescence study of the interaction of human apolipoprotein H with phospholipid vesicles.
Apolipoprotein H (ApoH) is a plasma glycoprotein with its in vivo physiological and pathogenic roles being closely related to its interaction with negatively charged membranes. In this paper, the interaction of ApoH with phospholipid vesicles was characterized by (i) detecting the wavelength shift of the fluorescence spectrum of ApoH and (ii) measuring the fluorescence quenching extent of ApoH by the membrane resident quencher 1-palmitoyl-2-stearoyl-(5-doxyl)-sn-glycero-3-phosphocholine (DPC). The observed blue shift upon addition of DMPG vesicles indicated that the tryptophan residues of ApoH moved from a polar to a nonpolar environment. The insertion ability of ApoH into PG-containing vesicles did not depend on the PG content in a stoichiometric way as did the blue shift, indicating that the negatively charged DMPG does not serve as a specific binding site but rather provides a suitable microenvironment for ApoH interaction. The finding that the detachment effect of cations on the blue shift is remarkably different from that on the quenching extent suggests that ApoH is capable of existing in two different conformations when membrane-bound.
State-Key Laboratory of Biomembrane, Department of Biological Sciences & Biotechnology, Tsinghua University, Beijing, People's Republic of China.
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D001053:Apolipoproteins / Q000737:chemistry / Q000378:metabolism*; D002118:Calcium / Q000737:chemistry; D002949:Citraconic Anhydrides / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D006358:Hot Temperature; D006801:Humans; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism*; D008956:Models, Chemical; D009994:Osmolar Concentration; D010084:Oxidation-Reduction; D010715:Phosphatidylglycerols / Q000737:chemistry / Q000378:metabolism; D010743:Phospholipids / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D013050:Spectrometry, Fluorescence; D014364:Tryptophan / Q000737:chemistry / Q000378:metabolism; D053482:beta 2-Glycoprotein I
D001053:Apolipoproteins; D002949:Citraconic Anhydrides; D004220:Disulfides; D006023:Glycoproteins; D008081:Liposomes; D010715:Phosphatidylglycerols; D010743:Phospholipids; D053482:beta 2-Glycoprotein I; D014364:Tryptophan; C002773:dimyristoylphosphatidylglycerol; D002118:Calcium