Channel activity of a viral transmembrane peptide in micro-BLMs: Vpu(1-32) from HIV-1.
We report for the first time on pore-suspending lipid bilayers, which we call micro-black lipid membranes (micro-BLMs), based on a highly ordered macroporous silicon array. Micro-BLMs were established by first functionalizing the backside porous silicon surface with gold and then chemisorbing 1,2-dipalmitoyl-sn-glycero-3-phosphothioethanol followed by spreading 1,2-diphytanoyl-sn-glycero-3-phosphocholine dissolved in n-decane. Impedance spectroscopy revealed the formation of single lipid bilayers confirmed by a mean specific capacitance of 0.6 +/- 0.2 microF/cm2. Membrane resistances were in the G omega-regime and beyond. The potential of the system for single channel recordings was demonstrated by inserting the transmembrane domain of the HIV-1 accessory peptide Vpu(1-32), which forms helix bundles with characteristic opening states. We elucidated different amilorides as potential drugs to inhibit channel activity of Vpu.
Römer
Winfried
W
Institut für Analytische Chemie, Chemo- und Biosensorik, Universität Regensburg, 93040 Regensburg, Germany.
Lam
Yuen H
YH
Fischer
Dahlia
D
Watts
Anthony
A
Fischer
Wolfgang B
WB
Göring
Petra
P
Wehrspohn
Ralf B
RB
Gösele
Ulrich
U
Steinem
Claudia
C
D000584:Amiloride / Q000031:analogs & derivatives / Q000494:pharmacology; D017097:Electric Impedance; D054298:Human Immunodeficiency Virus Proteins; D007473:Ion Channels / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D008051:Lipid Bilayers / Q000737:chemistry* / Q000378:metabolism; D008564:Membrane Potentials; D008567:Membranes, Artificial; D008855:Microscopy, Electron, Scanning; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D017434:Protein Structure, Tertiary; D012825:Silicon / Q000737:chemistry; D054334:Viral Regulatory and Accessory Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D000086082:Viroporin Proteins
D000584:Amiloride; D054298:Human Immunodeficiency Virus Proteins; D007473:Ion Channels; D008051:Lipid Bilayers; D008567:Membranes, Artificial; D010446:Peptide Fragments; D012825:Silicon; D054334:Viral Regulatory and Accessory Proteins; C514859:vpu protein, Human immunodeficiency virus 1; D000086082:Viroporin Proteins