Effect of phospholipids on the structure of Triatoma infestans lipophorin studied by fluorescence methods.
To study the role of phospholipids in the lipophorin structure, they were removed by phospholipase A2 treatment. Fluorescence lifetimes and accessibility to acrylamide quenching of different diphenylhexatrienyl derivatives, which were used as analogues of the different lipid components, indicate a surface localization of phospholipids and a distribution of diacylglycerols between the core and the surface, the surface fraction being increased by the phospholipase A2 treatment. The rotational behavior of these probes, studied by differential polarized phase fluorescence, indicates a high lipid order not only in the surface layer where phospholipids are located, but also in the core occupied by diacylglycerols and hydrocarbons. Phospholipid depletion increases the order only in the external region of the surface layer. Energy transfer from apolipoprotein tryptophan residues to several fluorescent acceptors indicates that phospholipid depletion produces a re-accommodation of the apoproteins. A decreased mobility of the water in the interfacial region is also produced by the phospholipase A2 treatment as it is reported by the fluorescence of 6-lauroyl-2-dimethylaminonaphthalene. This work shows that phospholipase A2 treatment of T. infestans lipophorin results in stable particles with an increased diacylglycerol to phospholipid ratio in the surface lipid layer. The modified particles are possibly stabilized by a conformational change in the apolipoproteins.
Instituto de Investigaciones Biocuímicas de la Plata (INIBIOLP), CONICET-UNLP, Facultad dé, Ciencias Médicas, Argentina.
O J
OJ
Rimoldi
H A
HA
Garda
R R
RR
Brenner
D015081:2-Naphthylamine / Q000031:analogs & derivatives / Q000737:chemistry; D000178:Acrylamides; D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D004161:Diphenylhexatriene; D004735:Energy Transfer; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes; D007848:Laurates / Q000737:chemistry; D008074:Lipoproteins / Q000737:chemistry* / Q000378:metabolism; D010741:Phospholipases A / Q000378:metabolism; D054467:Phospholipases A2; D010743:Phospholipids / Q000737:chemistry* / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D014224:Triatoma / Q000737:chemistry*; D014364:Tryptophan / Q000737:chemistry
D000178:Acrylamides; D002352:Carrier Proteins; D005456:Fluorescent Dyes; D007848:Laurates; D008074:Lipoproteins; D010743:Phospholipids; C032830:lipophorin; D004161:Diphenylhexatriene; D014364:Tryptophan; D015081:2-Naphthylamine; D010741:Phospholipases A; D054467:Phospholipases A2; C065580:laurdan