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13 Oct. 2005GW Canters Leiden Univ1. Scanning Probe Microscopy (SPM) Atomic/scanning Force...

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13 Oct. 2005 GW Canters Leiden Univ 1 III. Im aging A . S PM B. Fluorescent labeling
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Page 1: 13 Oct. 2005GW Canters Leiden Univ1. Scanning Probe Microscopy (SPM) Atomic/scanning Force Microscopy Scanning Tunnelling Microscopy.

13 Oct. 2005 GW Canters Leiden Univ 1

I I I . Imaging

A. SPM

B. Fluorescent labeling

Page 2: 13 Oct. 2005GW Canters Leiden Univ1. Scanning Probe Microscopy (SPM) Atomic/scanning Force Microscopy Scanning Tunnelling Microscopy.

Scanning Probe Microscopy (SPM)

Atomic/scanning Force Microscopy

Scanning Tunnelling Microscopy

Page 3: 13 Oct. 2005GW Canters Leiden Univ1. Scanning Probe Microscopy (SPM) Atomic/scanning Force Microscopy Scanning Tunnelling Microscopy.

AFM resolution

Chris Chan, MetProt Group, private comm. 2004

Page 4: 13 Oct. 2005GW Canters Leiden Univ1. Scanning Probe Microscopy (SPM) Atomic/scanning Force Microscopy Scanning Tunnelling Microscopy.

Mica 5nm Au nanoparticle

Chris Chan, MetProt Group, private comm. 2004

Page 5: 13 Oct. 2005GW Canters Leiden Univ1. Scanning Probe Microscopy (SPM) Atomic/scanning Force Microscopy Scanning Tunnelling Microscopy.

2 & 5nm Au particles on mica, TM in water

Chris Chan, MetProt Group, private comm. 2004

Page 6: 13 Oct. 2005GW Canters Leiden Univ1. Scanning Probe Microscopy (SPM) Atomic/scanning Force Microscopy Scanning Tunnelling Microscopy.

5nm Au particles on HOPG

Chris Chan, MetProt Group, private comm. 2004

Page 7: 13 Oct. 2005GW Canters Leiden Univ1. Scanning Probe Microscopy (SPM) Atomic/scanning Force Microscopy Scanning Tunnelling Microscopy.

13 Oct. 2005 GW Canters Leiden Univ 7

J.Davis et al., Chem Communic. (2003) Advance pub.

Page 8: 13 Oct. 2005GW Canters Leiden Univ1. Scanning Probe Microscopy (SPM) Atomic/scanning Force Microscopy Scanning Tunnelling Microscopy.

13 Oct. 2005 GW Canters Leiden Univ 8

J.Davis Pers. Communic. (2003)

Page 9: 13 Oct. 2005GW Canters Leiden Univ1. Scanning Probe Microscopy (SPM) Atomic/scanning Force Microscopy Scanning Tunnelling Microscopy.

13 Oct. 2005 GW Canters Leiden Univ 9

Forcing current through a single metalloprotein

Fluid conductance

J.Davis Pers. Communic. (2003)

Page 10: 13 Oct. 2005GW Canters Leiden Univ1. Scanning Probe Microscopy (SPM) Atomic/scanning Force Microscopy Scanning Tunnelling Microscopy.

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?

Page 11: 13 Oct. 2005GW Canters Leiden Univ1. Scanning Probe Microscopy (SPM) Atomic/scanning Force Microscopy Scanning Tunnelling Microscopy.

13 Oct. 2005 GW Canters Leiden Univ 11

Flavodoxin

Page 12: 13 Oct. 2005GW Canters Leiden Univ1. Scanning Probe Microscopy (SPM) Atomic/scanning Force Microscopy Scanning Tunnelling Microscopy.

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

Page 13: 13 Oct. 2005GW Canters Leiden Univ1. Scanning Probe Microscopy (SPM) Atomic/scanning Force Microscopy Scanning Tunnelling Microscopy.

The trimeric structure of copper-containing NiR

Monomer Trimer

Page 14: 13 Oct. 2005GW Canters Leiden Univ1. Scanning Probe Microscopy (SPM) Atomic/scanning Force Microscopy Scanning Tunnelling Microscopy.

Nir on template stripped gold10ug/ml in 20mM MES, pH 6TM in air

Chris Chan, MetProt Group, private comm. 2004

Page 15: 13 Oct. 2005GW Canters Leiden Univ1. Scanning Probe Microscopy (SPM) Atomic/scanning Force Microscopy Scanning Tunnelling Microscopy.

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

Page 16: 13 Oct. 2005GW Canters Leiden Univ1. Scanning Probe Microscopy (SPM) Atomic/scanning Force Microscopy Scanning Tunnelling Microscopy.

1ug/ml Nir M94C, polylysine on mica

TM in 20mM MES, pH6Chris Chan, MetProt Group, private comm. 2004

Page 17: 13 Oct. 2005GW Canters Leiden Univ1. Scanning Probe Microscopy (SPM) Atomic/scanning Force Microscopy Scanning Tunnelling Microscopy.

13 Oct. 2005 GW Canters Leiden Univ 17

I I I . Imaging

A. SPM

B. Fluorescent labeling

Page 18: 13 Oct. 2005GW Canters Leiden Univ1. Scanning Probe Microscopy (SPM) Atomic/scanning Force Microscopy Scanning Tunnelling Microscopy.

13 Oct. 2005 GW Canters Leiden Univ 18Nature Reviews Mol. Cell Biol. 3(2002)906

JACS 124(2002)6063

PNAS 101(2004)523

Page 19: 13 Oct. 2005GW Canters Leiden Univ1. Scanning Probe Microscopy (SPM) Atomic/scanning Force Microscopy Scanning Tunnelling Microscopy.

13 Oct. 2005 GW Canters Leiden Univ 19

Nature Reviews Mol. Cell Biol. 3(2002)906


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