Covalent anchoring of proteins onto gold-directed NHS-terminated self-assembled monolayers in aqueous buffers: SFM images of clathrin cages and triskelia.
N-Hydroxysuccinimide-terminated self-assembled monolayers with linear (CH2)10 chains were prepared on ultraflat Au(111) surfaces from dithiobis(succinimidylundecanoate). These monolayers, which are covalently chemisorbed to gold via thiolate bonds, form a highly reactive amino-group specific carpet at the liquid-solid interface. Proteins bind to it covalently in aqueous buffers under mild conditions; this provides a (general) procedure for protein immobilization for scanning probe microscopy. Using this technique, we have obtained what we believe are the first scanning force microscopy images of clathrin cages and of their in situ disassembly, yielding typical triskelia under non-denaturing conditions.
Wagner
P
P
Department of Biochemistry, Swiss Federal Institute of Technology, Zurich.
Kernen
P
P
Hegner
M
M
Ungewickell
E
E
Semenza
G
G
D000818:Animals; D001921:Brain; D002021:Buffers; D002417:Cattle; D002966:Clathrin / Q000648:ultrastructure*; D003034:Coated Pits, Cell-Membrane / Q000737:chemistry; D006046:Gold; D007202:Indicators and Reagents; D018625:Microscopy, Atomic Force / Q000379:methods; D013388:Succinimides
D002021:Buffers; D002966:Clathrin; D007202:Indicators and Reagents; D013388:Succinimides; D006046:Gold; C001426:N-hydroxysuccinimide