Movatterモバイル変換

[0]ホーム
URL:

Skip to main content
Springer Nature Link
Log in

1H NMR studies of deuterated ribonuclease HI selectively labeled with protonated amino acids

  • Research Papers
  • Published:
Journal of Biomolecular NMR Aims and scope Submit manuscript

Summary

Two-dimensional (2D)1H NMR experiments using deuterium labeling have been carried out to investigate the solution structure of ribonuclease HI (RNase HI) fromEscherichia coli (E. coli), which consists of 155 amino acids. To simplify the1H NMR spectra, two fully deuterated enzymes bearing several prototed amino acids were prepared from an RNase HI overproducing strain ofE. coli grown in an almost fully deuterated medium. One enzyme was selectively labeled by protonated His, He. Val. and Leu. The other was labeled by only protonated His and Ile. The 2D1H NMR spectra of these deuterated R Nase H1 proteins, selectively labeled with protonated amino acids, were much more simple than those of the normally protonated enzyme. The simplified spectra allowed unambiguous assignments of the resonance peaks and connectivities in COSY and NOESY for the side-chain protons. The spin-lattice relaxation times of the side-chain protons of the buried His residue of the deuterated enzyme became remarkably longer than that of the protonated enzyme. In contrast, the relaxation times of the side-chain protons of exposed His residues remained essentially unchanged.

This is a preview of subscription content,log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+
from ¥17,985 /Month
  • Starting from 10 chapters or articles per month
  • Access and download chapters and articles from more than 300k books and 2,500 journals
  • Cancel anytime
View plans

Buy Now

Price includes VAT (Japan)

Instant access to the full article PDF.

Similar content being viewed by others

ArticleOpen access14 February 2019

Explore related subjects

Discover the latest articles, books and news in related subjects, suggested using machine learning.

References

  • Atkinson, J.G., Csakvary, J.J., Herbert, G.T. and Stuart, R.S. (1968)J. Am. Chem. Soc.,90, 498–499.

    Google Scholar 

  • Brodin, P., Drakenberg, T., Thulin, E., Forsén, S. and Grundström, T. (1989)Prot. Eng.,2, 353–358.

    Google Scholar 

  • Crespi, H.L., Rosenberg, R.M. and Katz, J.J. (1968)Science,161, 795–796.

    Google Scholar 

  • Crouch, R.J. and Dirksen, M.-L. (1982) InNuclease (Eds Linn, S.M. and Roberts, R.J.) Cold Spring Harbor Laboratory. Cold Spring Harbor, NY, pp. 211–241.

    Google Scholar 

  • Doolittle, R.F., Feng, D.-F., Johnson, M.S. and McClure, M.A. (1989)Q. Rev. Biol.,64, 1–30.

    Google Scholar 

  • Fesik, S.W. and Zuiderweg, E.R.P. (1990)Q. Rev. Biophys.,23, 97–131.

    Google Scholar 

  • Ikura, M., Kay, L.E. and Bax, A. (1990)Biochemistry,29, 4659–4667.

    Google Scholar 

  • Itaya, M. (1990)Proc. Natl. Acad. Sci. U.S.A.,87: 8587–8591.

    Google Scholar 

  • Johnson, M.S., McClure, M.A., Feng, D.-F., Gray, J. and Doolittle, R.F. (1986)Proc. Natl. Acad. Sci. U.S.A.,83, 7648–7652.

    Google Scholar 

  • Kanaya, S. and Crouch, R.J. (1983)J. Biol. Chem.,258, 1276–1281.

    Google Scholar 

  • Kanaya, S., Kohara, A., Miyagawa, M., Matsuzaki, T., Morikawa, K. and Ikehara, M. (1989)J. Biol. Chem.,264, 11546–11549.

    Google Scholar 

  • Kanaya, S., Kohara, A., Miura, Y., Sekiguchi, A., Iwai, S., Inoue, H., Ohtsuka, E. and Ikehara, M. (1990)J. Biol. Chem.,264, 4615–4621.

    Google Scholar 

  • Katayanagi, K., Miyagawa, M., Matsushima, M., Ishikawa, M., Kanaya, S., Ikehara, M., Matsuzaki, T. and Morikawa, K. (1990)Nature,347, 306–309.

    Google Scholar 

  • Katayanagi, K., Miyagawa, M., Matsushima, M., Ishikawa, M., Kanaya, S., Nakamura, H., Ikehara, M., Matsuzaki, T. and Morikawa, K. (1991)J. Mol. Biol., submitted.

  • Kay, L.E., Clore, G.M., Bax, A. and Gronenborn, A.M. (1990)Science,249, 411–414.

    Google Scholar 

  • LeMaster, D.M. (1990)Q. Rev. Biophys.,23, 133–174.

    Google Scholar 

  • Marion, D. and Wüthrich, K. (1983)Biochem Biophys. Res. Commun.,113, 967–974.

    Google Scholar 

  • Markley, J.L., Putter, I. and Jardetzky, O. (1968)Science,161, 1249–1251.

    Google Scholar 

  • McIntosh, L.P. and Dahlquist, F.W. (1990)Q. Rev. Biophys.,23, 1–38.

    Google Scholar 

  • Nagayama, K., Yamazaki, T., Yoshida, M., Kanaya, S. and Nakamura, H. (1990)J. Biochem.,108, 149–152.

    Google Scholar 

  • Nagayama, K., Yamazaki, T., Yoshida, M., Kanaya, S. and Nakamura, H. (1991) InComputational Aspects of the Study of the Biological Macromolecules by NMR (NATO. ASI series) (Ed, Hoch, J.C.), in press.

  • Rodwell, V.W. (1969) InMetabolic Pathways. Vol. 3 (Ed Greenberg, D.M.) Academic Press, New York, p. 351.

    Google Scholar 

  • Seeholzer, S.H., Cohn, M., Putkey, J.A., Means, A.R. and Crespi, H.L. (1986)Proc. Natl. Acad. Sci. U.S.A.,83, 3634–3638.

    Google Scholar 

  • Shrake, A. and Rupley, J.A. (1973)J. Mol. Biol.,79, 351–371.

    Google Scholar 

  • Takahashi, S. and Nagayama, K. (1988)J. Magn. Reson.,76, 347–351.

    Google Scholar 

  • Wüthrich, K. (1986)NMR of Proteins and Nucleic Acids, John Wiley and Sons, New York.

    Google Scholar 

  • Yamazaki, T., Yoshida, M., Kanaya, S., Nakamura, H. and Nagayama, K. (1991)Biochemistry,30, 6036–6047.

    Google Scholar 

  • Yang, W., Hendrickson, W.A., Crouch, R.J. and Satow, Y. (1990)Science249, 1398–1405.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Protein Engineering Research Institute, 6-2-3 Furuedai, 565, Suita Osaka, Japan

    Yasushi Oda, Haruki Nakamura, Shigenori Kanaya & Morio Ikehara

  2. Biometrology Laboratory, JEOL Ltd., Akishima, 196, Tokyo, Japan

    Toshio Yamazaki & Kuniaki Nagayama

  3. Tokyo Research Laboratories, Kyowa Hakko Kogyo Co. Ltd., Machida, 194, Tokyo, Japan

    Mayumi Yoshida

Authors
  1. Yasushi Oda
  2. Haruki Nakamura
  3. Toshio Yamazaki
  4. Kuniaki Nagayama
  5. Mayumi Yoshida
  6. Shigenori Kanaya
  7. Morio Ikehara

Rights and permissions

About this article

Cite this article

Oda, Y., Nakamura, H., Yamazaki, T.et al.1H NMR studies of deuterated ribonuclease HI selectively labeled with protonated amino acids.J Biomol NMR2, 137–147 (1992). https://doi.org/10.1007/BF01875525

Download citation

Keywords

Access this article

Subscribe and save

Springer+
from ¥17,985 /Month
  • Starting from 10 chapters or articles per month
  • Access and download chapters and articles from more than 300k books and 2,500 journals
  • Cancel anytime
View plans

Buy Now

Price includes VAT (Japan)

Instant access to the full article PDF.

Advertisement

[8]ページ先頭
©2009-2025 Movatter.jp