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Crystal structure of rubredoxin from Pyrococcus furiosus at 0.95 Å resolution, and the structures of N-terminal methionine and formylmethionine variants of Pf Rd. Contributions of N-terminal interactions to thermostability

Bau, Robert and Rees, Douglas C. and Kurtz, Donald M., Jr. and Scott, Robert A. and Huang, Heshu and Adams, Michael W. W. and Eidsness, Marly K. (1998) Crystal structure of rubredoxin from Pyrococcus furiosus at 0.95 Å resolution, and the structures of N-terminal methionine and formylmethionine variants of Pf Rd. Contributions of N-terminal interactions to thermostability. Journal of Biological Inorganic Chemistry, 3 (5). pp. 484-493. ISSN 0949-8257. https://resolver.caltech.edu/CaltechAUTHORS:20150108-160800902

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Abstract

The high-resolution crystal structure of the small iron-sulfur protein rubredoxin (Rd) from the hyperthermophilic archeon Pyrococcus furiosus (Pf) is reported in this paper, together with those of its methionine ([_0M]Pf Rd) and formylmethionine (f[_0M]Pf Rd) variants. These studies were conducted to assess the consequences of the presence or absence of a salt bridge between the amino terminal nitrogen of Ala1 and the side chain of Glu14 to the structure and stability of this rubredoxin. The structure of wild-type Pf Rd was solved to a resolution of 0.95 Å and refined by full-matrix least-squares techniques to a crystallographic agreement factor of 12.8% [F>2σ(F) data, 25 617 reflections], while those of the [_0M]Pf and f[_0M]Pf Rd variants were solved at slightly lower resolutions (1.1 Å, R=11.5%, 17 213 reflections; 1.2 Å, R=13.7%, 12 478 reflections, respectively). The quality of the data was such that about half of the hydrogen atoms of the protein were clearly visible. All three structures were ultimately refined using the program SHELXL-93 with anisotropic atomic displacement parameters for all non-hydrogen protein atoms, and calculated hydrogen positions included but not refined. In this paper we also report thermostability data for all three forms of Pf Rd, and show that they follow the sequence wild-type >[_0M]Pf>formyl[_0M]Pf. Comparison of the three Pf Rd structures in the N-terminal region show that the structures of wild-type Pf Rd and f[_0M]Pf are rather similar, while that of [_0M]Pf Rd shows a number of additional hydrogen bonds involving the extra methionine group. While the salt bridge between the Ala1 amino group and the Glu14 carboxylate is not the primary determinant of the thermostability of Pf Rd, alterations to the amino terminus do have a moderate influence on the thermostability of this protein.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1007/s007750050258 DOIArticle
http://link.springer.com/article/10.1007%2Fs007750050258PublisherArticle
http://rdcu.be/pY7JPublisherFree ReadCube access
ORCID:
AuthorORCID
Rees, Douglas C.0000-0003-4073-1185
Additional Information:© 1998 SBIC. Received: 27 March 1998; Accepted: 25 June 1998. This work was supported by the National Science Foundation (Grant DMB 91-18689 to D.C.R., Grant BCS-96-32657 to M.W.W.A.), the National Institutes of Health (Grant GM-50736 to D.M.K.), and NSF Research Training Group Award DIR-90-14281 to the Center for Metalloenzyme Studies, University of Georgia. R.B. acknowledges sabbatical leave support from the University of Southern California.
Funders:
Funding AgencyGrant Number
NSFDMB 91-18689
NSFBCS-96-32657
NIHGM-50736
NSFDIR-90-14281
Subject Keywords:rubredoxin; iron-sulfur proteins; hyperthermostability; protein structure; metalloproteins
Issue or Number:5
Record Number:CaltechAUTHORS:20150108-160800902
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20150108-160800902
Official Citation:Bau, R., Rees, D., Kurtz Jr., D. et al. JBIC (1998) 3: 484. doi:10.1007/s007750050258
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:53429
Collection:CaltechAUTHORS
Deposited By: SWORD User
Deposited On:13 Jan 2015 15:54
Last Modified:03 Oct 2019 07:49

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