Biomedical subjects

A De Lisi

Publications and source records attributed to A De Lisi.

4 recordsLinked to original sources

PubMed1995

EPR study of annexin V-cardiolipin Ca-mediated interaction in phospholipid vesicles and isolated mitochondria.

The properties of the binding of annexin V to variously composed phospholipid vesicles have been studied by applying a recently developed EPR method, using an annexin V spin label. By this approach, this protein is seen to bind to acidic phospholipid-containing vesicles, as reported, thus confirming the reliability of the method. In addition, binding of this annexin to cardiolipin-containing vesicles has been studied in more depth, and the protein has been shown to have a distinct affinity for this phospholipid. As a cardiolipin-rich natural membrane system, mitochondrial membranes and mitoplasts from rat liver were considered, and a strong binding of AV to these membranes was observed. Having compared this binding with that to phospholipid vesicles, cardiolipin-rich microdomains in the mitochondrial membranes are proposed as the putative mitochondrial binding sites for annexin V.

Animals↗

PubMed1993

An EPR method for studying annexin-biomembrane interaction.

Annexin p34 spin label has been used as a probe for studying annexin-membrane Ca-mediated interaction. The EPR spectrum of the purified probe is able to monitor the binding of the protein to phospholipid vesicles and isolated mitochondrial membranes, with increasing calcium concentration, as revealed by the calculated rotational correlation time. A novel mathematical approach is proposed to provide a direct estimation of annexin percentage binding to membranes, based on the correlation time calculated from experimental EPR spectra.

Animals↗

PubMed1990

Spin labeling of calcium-dependent phospholipid-binding proteins.

Bovine lung annexins p32 and p34 were spin labeled with an iodoacetamidoproxyl spin label, a reagent that reportedly couples with protein methionine residues. Labeling conditions and stoichiometry were studied with the radiolabeled analogue [1-14C]iodoacetamide. As judged by this method, carboxamidomethylation of both p32 and p34 occurred up to a 0.7 mol ratio after 60 h of reaction at 37 degrees C and at pH 4. The two proteins retained Ca2(+)-dependent phospholipid-binding ability both in radiolabeled and in spin-labeled forms. Electron resonance spectra of spin-labeled p32 and p34 showed the features of a partially immobilized spin probe, with rotational correlation time values of 1.15 and 1.25 ns, respectively, which definitely indicate successful spin labeling. Quantitation of ESR spectra by computer double integration indicated 70% spin labeling of both proteins, as anticipated by radiolabeling. The use of spin-labeled p32 and p34 in the study of Ca2(+)-dependent interaction of annexins with biomembranes is proposed.

Animals↗

PubMed1986

Nonspecific lipid transfer protein (sterol carrier protein 2) is bound to rat liver mitochondria: its role in spontaneous intermembrane phospholipid transfer.

In the present study we have investigated the transfer of phospholipids between vesicles and rat liver mitochondria. Transfer was measured by electron paramagnetic resonance spectroscopy using vesicles that contained spin-labeled phospholipids. A spontaneous transfer was observed which could be strongly inhibited by treating the mitochondria with the thiol reagent mersalyl. Transfer was also greatly reduced after a saline wash of the mitochondria; the transfer activity was then recovered in the wash. This activity was inhibited by tryptic digestion and mersalyl. By gel chromatography, enzyme immunoassay and immunoblotting it was demonstrated that the activity in the wash was due to the nonspecific lipid transfer protein (sterol carrier protein 2). We could estimate that up to 85% of the spontaneous phospholipid transfer between vesicles and rat liver mitochondria was mediated by this transfer protein.

Animals↗