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13 Oct. 2005 GW Canters Leiden Univ 1
I I I . Imaging
A. SPM
B. Fluorescent labeling
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Scanning Probe Microscopy (SPM)
Atomic/scanning Force Microscopy
Scanning Tunnelling Microscopy
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AFM resolution
Chris Chan, MetProt Group, private comm. 2004
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Mica 5nm Au nanoparticle
Chris Chan, MetProt Group, private comm. 2004
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2 & 5nm Au particles on mica, TM in water
Chris Chan, MetProt Group, private comm. 2004
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5nm Au particles on HOPG
Chris Chan, MetProt Group, private comm. 2004
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13 Oct. 2005 GW Canters Leiden Univ 7
J.Davis et al., Chem Communic. (2003) Advance pub.
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13 Oct. 2005 GW Canters Leiden Univ 8
J.Davis Pers. Communic. (2003)
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13 Oct. 2005 GW Canters Leiden Univ 9
Forcing current through a single metalloprotein
Fluid conductance
J.Davis Pers. Communic. (2003)
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13 Oct. 2005 GW Canters Leiden Univ 10
J.D. Davis, D. Bruce, G.W. Canters, J. Crozier, H.A.O. Hill (2003) Chem. Commun. 576-577.
Height distribution was found to be the same for the wt and K27C azurins, while the height of S118C is 5 - 8 Å lower
AFM permits to distinguish between the “standing” and the “lying” azurin molecules.
Electrochemical characteristics of the azurin monolayers.
Half wave potential is defined as (Epa–Epc)/2 vs. SCE
Azurin WT Azurin K27C Azurin S118C
E1/2*(mV vs. SCE) 165–175 165–175 120–145
ket/s–1 300–400 210–320 300–570
BUT: the electron transfer rate is independent of the distance between the Cu and the electrode surface
Is it possibly an effect of heterogeneous binding?
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13 Oct. 2005 GW Canters Leiden Univ 11
Flavodoxin
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13 Oct. 2005 GW Canters Leiden Univ 12
Flavodoxin N106C dimer on mica~0.1ug/ml in 20mM MES pH6TM in air
STM images of tip-induced dissociation of K27C azurin dimer, (a) before and (b) after application of a voltage pulse. Scanning conditions were bias voltage 0.1 V, current 0.1 nA, temperature 20±1 °C, relative humidity 35±5%. Halliwell et.al. 2004
Chris Chan, MetProt Group, private comm. 2004
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The trimeric structure of copper-containing NiR
Monomer Trimer
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Nir on template stripped gold10ug/ml in 20mM MES, pH 6TM in air
Chris Chan, MetProt Group, private comm. 2004
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13 Oct. 2005 GW Canters Leiden Univ 15
Nir on template stripped gold
100ug/ml in 20mM MES, pH 6
TM in air
Chris Chan, MetProt Group, private comm. 2004
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1ug/ml Nir M94C, polylysine on mica
TM in 20mM MES, pH6Chris Chan, MetProt Group, private comm. 2004
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13 Oct. 2005 GW Canters Leiden Univ 17
I I I . Imaging
A. SPM
B. Fluorescent labeling
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13 Oct. 2005 GW Canters Leiden Univ 18Nature Reviews Mol. Cell Biol. 3(2002)906
JACS 124(2002)6063
PNAS 101(2004)523
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13 Oct. 2005 GW Canters Leiden Univ 19
Nature Reviews Mol. Cell Biol. 3(2002)906