Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 1842-1848 |
| Number of pages | 7 |
| Journal | Biophysical Journal |
| Volume | 72 |
| Issue number | 4 |
| Publication status | Published - 1997 Apr |
| Externally published | Yes |
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ASJC Scopus subject areas
- Biophysics
Cite this
Two-dimensional arrangement of a functional protein by cysteine-gold interaction : Enzyme activity and characterization of a protein monolayer on a gold substrate. / Sasaki, Yuji C.; Yasuda, Kenji; Suzuki, Yoshio; Ishibashi, Tadashi; Satoh, Isamu; Fujiki, Yasutake; Ishiwata, Shin'ichi.
In: Biophysical Journal, Vol. 72, No. 4, 04.1997, p. 1842-1848.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Two-dimensional arrangement of a functional protein by cysteine-gold interaction
T2 - Enzyme activity and characterization of a protein monolayer on a gold substrate
AU - Sasaki, Yuji C.
AU - Yasuda, Kenji
AU - Suzuki, Yoshio
AU - Ishibashi, Tadashi
AU - Satoh, Isamu
AU - Fujiki, Yasutake
AU - Ishiwata, Shin'ichi
PY - 1997/4
Y1 - 1997/4
N2 - 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.
AB - 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=0030939055&partnerID=8YFLogxK
M3 - Article
C2 - 9083688
AN - SCOPUS:0030939055
VL - 72
SP - 1842
EP - 1848
JO - Biophysical Journal
JF - Biophysical Journal
SN - 0006-3495
IS - 4
ER -