Evidence that small proteins translocate through silicon nitride pores in a folded conformation

• Published in: Journal of physics. Condensed matter Vol. 22; no. 45; p. 454133
• Main Authors: Stefureac, Radu I, Trivedi, Dhruti, Marziali, Andre, Lee, Jeremy S
• Format: Journal Article
• Language: English
• Published: Bristol IOP Publishing 17.11.2010; Institute of Physics
• Subjects: Analytical, structural and metabolic biochemistry; Biological and medical sciences; Diffusion; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; Motion; Nanostructures - chemistry; Nanostructures - ultrastructure; Particle Size; Porosity; Protein Conformation; Protein Folding; Proteins; Proteins - chemistry; Silicon Compounds - chemistry; Translocation; Electrophoresis; Nanopore; Hydration; Protein
• ISSN: 0953-8984; 1361-648X
• DOI: 10.1088/0953-8984/22/45/454133

The interaction of three proteins (histidine-containing phosphocarrier protein, HPr, calmodulin, CaM, and maltose binding protein, MBP) with synthetic silicon nitride (SiN(x)) membranes has been studied. The proteins which have a net negative charge were electrophoretically driven into pores of 7 and 5 nm diameter with a nominal length of 15 nm. The % blockade current and event duration were measured at three different voltages. For a translocation event it was expected that the % block would be constant with voltage whilst the event duration would decrease with increasing voltage. On the basis of these criteria, we deduce that MBP whose largest dimension is 6.5 nm does not translocate whereas up to 40% of CaM molecules can translocate the 7 nm pore as can a majority of HPr molecules, with some translocations being observed for the 5 nm pore. For translocation events the magnitude of the % blockade current is consistent with a folded conformation of the proteins surrounded by a hydration shell of 0.5-1.0 nm.