Mechanism of mitochondrial membrane permeabilization by HIV-1 Vpr, mimetics of Vpr and methods of screening active molecules having the ability to alter and/or prevent and/or mimic the interaction of Vpr with ANT

A method of screening for molecules that compete with the binding of the C-terminal moeity of Vpr to ANT comprising:

• (a) providing a Vpr fragment capable of binding to ANT;
• (b) contacting said Vpr fragment with an ANT fragment capable of binding to Vpr in the presence and absence of a test molecule;
• and
• (c) detecting the binding of said Vpr fragment to said ANT fragment in the presence and absence of a test molecule.

A method of screening for molecules that mimic Vpr or Vpr fragments in its capacity to interact physically of with ANT comprising:

• a) providing a Vpr or Vpr fragment capable of interacting with ANT,
• b) contacting said Vpr or Vpr fragment with an ANT fragment capable of interacting with Vpr or Vpr fragment in the presence of absence of a test molecule;
• and
• c) detecting the binding of said Vpr or Vpr fragment to said ANT fragment in the presence of absence of a test molecule.

A method of screening for genetic or epigenetic alterations in the expression or structure of the three ANT isoforms in humans comprising:

• (a) providing a fragment of Vpr, wherein said fragment is capable of binding to ANT, with a sample comprising human ANT;
• (b) mixing said fragment with a biological sample comprising human ANT;
• (c) mixing said fragment with a control sample comprising human ANT;
• (d) detecting the binding of Vpr to ANT in said biological sample and said control sample;
• (e) correlating a difference in binding with a genetic or epigenetic alteration of ANT;
• and
• (f) optionally detecting a difference in the ANT capacity to form channel in liposome or in planar lipids bilayers.

A method of screening active molecules of interest that induce or prevent formation of a lethal pore by ANT comprising:

• (a) providing purified ANT in artificial lipid bilayers or liposomes;
• (b) contacting molecules of interest to be screened with said ANT;
• and
• (c) detecting lethal pore formation by measuring the release of labeled substrate.

A method of screening active molecules of interest that inhibit the formation of a lethal pore without preventing antiport function comprising:

• (a) providing a composition comprising purified ANT in artificial lipid bilayers or liposomes with a molecule that induces the formation of a lethal pore;
• (b) contacting said composition in the presence or absence of a test molecule.
• (c) detecting by fluorescence the presence of the antiport function;
• and
• (d) detecting by another fluorescence the test molecule that inhibits the formation of a lethal pore.