PMID- 25670233
OWN - NLM
STAT- MEDLINE
DCOM- 20151124
LR  - 20150303
IS  - 1520-5827 (Electronic)
IS  - 0743-7463 (Linking)
VI  - 31
IP  - 8
DP  - 2015 Mar 3
TI  - Silica nanoparticles for the oriented encapsulation of membrane proteins into 
      artificial bilayer lipid membranes.
PG  - 2511-6
LID - 10.1021/la504417j [doi]
AB  - An artificial bilayer lipid membrane system is presented, featuring the oriented 
      encapsulation of membrane proteins in a functionally active form. Nickel 
      nitrilo-triacetic acid-functionalized silica nanoparticles, of a diameter of 
      around 25 nm, are used to attach the proteins via a genetically engineered 
      histidine tag in a uniform orientation. Subsequently, the proteins are 
      reconstituted within a phospholipid bilayer, formed around the particles by in 
      situ dialysis to form so-called proteo-lipobeads (PLBs). With a final size of 
      about 50 nm, the PLBs can be employed for UV/vis spectroscopy studies, 
      particularly of multiredox center proteins, because the effects of light 
      scattering are negligible. As a proof of concept, we use cytochrome c oxidase 
      (CcO) from P. denitrificans with the his tag genetically engineered to subunit I. 
      In this orientation, the P side of CcO is directed to the outside and hence 
      electron transfer can be initiated by reduced cytochrome c (cc). UV/vis 
      measurements are used in order to determine the occupancy by CcO molecules 
      encapsulated in the lipid bilayer as well as the kinetics of electron transfer 
      between CcO and cc. The kinetic data are analyzed in terms of the 
      Michaelis-Menten kinetics showing that the turnover rate of CcO is significantly 
      decreased compared to that of solubilized protein, whereas the binding 
      characteristics are improved. The data demonstrate the suitability of PLBs for 
      functional cell-free bioassays of membrane proteins.
FAU - Schadauer, Florian
AU  - Schadauer F
AD  - Austrian Institute of Technology GmbH , AIT, Donau-City Str. 1, 1220 Vienna, 
      Austria.
FAU - Geiss, Andreas F
AU  - Geiss AF
FAU - Srajer, Johannes
AU  - Srajer J
FAU - Siebenhofer, Bernhard
AU  - Siebenhofer B
FAU - Frank, Pinar
AU  - Frank P
FAU - Reiner-Rozman, Ciril
AU  - Reiner-Rozman C
FAU - Ludwig, Bernd
AU  - Ludwig B
FAU - Richter, Oliver-M H
AU  - Richter OM
FAU - Nowak, Christoph
AU  - Nowak C
FAU - Naumann, Renate L C
AU  - Naumann RL
LA  - eng
PT  - Journal Article
DEP - 20150220
PL  - United States
TA  - Langmuir
JT  - Langmuir : the ACS journal of surfaces and colloids
JID - 9882736
RN  - 0 (Lipid Bilayers)
RN  - 0 (Membrane Proteins)
RN  - 0 (Phospholipids)
RN  - 7631-86-9 (Silicon Dioxide)
RN  - EC 1.9.3.1 (Electron Transport Complex IV)
SB  - IM
MH  - Electron Transport Complex IV/*chemistry/metabolism
MH  - Lipid Bilayers/chemical synthesis/*chemistry
MH  - Membrane Proteins/*chemistry
MH  - Models, Molecular
MH  - Nanoparticles/*chemistry
MH  - Paracoccus denitrificans/enzymology
MH  - Particle Size
MH  - Phospholipids/chemical synthesis/*chemistry
MH  - Silicon Dioxide/*chemistry
MH  - Surface Properties
EDAT- 2015/02/12 06:00
MHDA- 2015/12/15 06:00
CRDT- 2015/02/12 06:00
PHST- 2015/02/12 06:00 [entrez]
PHST- 2015/02/12 06:00 [pubmed]
PHST- 2015/12/15 06:00 [medline]
AID - 10.1021/la504417j [doi]
PST - ppublish
SO  - Langmuir. 2015 Mar 3;31(8):2511-6. doi: 10.1021/la504417j. Epub 2015 Feb 20.