Two-dimensional arrangement of a functional protein by cysteine-gold interaction: enzyme activity and characterization of a protein monolayer on a gold substrate.
We have characterized the functional protein, myosin subfragment 1 (S1), attached to a gold substrate by the sulfhydryl groups of cysteine in proteins. The amino groups of the regulatory light chain (RLC) isolated from myosin were labeled with a radioisotope (125I), and the labeled RLC was incorporated into S1 from which the RLC had been removed. The radiation from 125I showed that S1 molecules had attached to the gold and, through the interference effect of the monochromatic radiation from 125I, provided information about the position of labeled RLC sites in the S1 monolayer. The interference fringes showed that the RLC was located close to the gold surface and that all of the adsorbed S1 molecules had the same orientation. We confirmed that the motor function of S1 on the gold surface is maintained by observing sliding movement at low ionic strength and by observing the detachment at high ionic strength of fluorescent actin filaments in the presence of ATP. We also found that the adsorbed S1 molecules were not removed from the Au surface by a reducing agent. Thus the Au-S bond is more stable than the S-S bond.
Sasaki
Y C
YC
Advanced Research Laboratory, Hitachi Ltd., Saitama, Japan. ycsasaki@harl.hitachi.co.jp
Yasuda
K
K
Suzuki
Y
Y
Ishibashi
T
T
Satoh
I
I
Fujiki
Y
Y
Ishiwata
S
S
D000199:Actins / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000327:Adsorption; D003545:Cysteine / Q000378:metabolism; D005456:Fluorescent Dyes; D006046:Gold / Q000378:metabolism*; D007457:Iodine Radioisotopes; D008958:Models, Molecular; D018994:Myosin Light Chains; D015879:Myosin Subfragments / Q000737:chemistry* / Q000378:metabolism; D010590:Phalloidine / Q000378:metabolism; D011487:Protein Conformation; D012235:Rhodamines / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000378:metabolism; D013499:Surface Properties; D014965:X-Rays
D000199:Actins; D005456:Fluorescent Dyes; D007457:Iodine Radioisotopes; D018994:Myosin Light Chains; D015879:Myosin Subfragments; D012235:Rhodamines; D013438:Sulfhydryl Compounds; D010590:Phalloidine; D006046:Gold; D000255:Adenosine Triphosphate; D003545:Cysteine