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PMID: 2184035 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Selenomethionyl proteins produced for analysis by multiwavelength anomalous diffraction (MAD): a vehicle for direct determination of three-dimensional structure.

The EMBO journal ·Vol. 9 ·No. 5 ·1990-05-00 ·Pages 1665-72

Hendrickson WA, Horton JR, LeMaster DM

Abstract

An expression system has been established for the incorporation of selenomethionine into recombinant proteins produced from plasmids in Escherichia coli. Replacement of methionine by selenomethionine is demonstrated at the level of 100% for both T4 and E. coli thioredoxins. The natural recombinant proteins and the selenomethionyl variants of both thioredoxins crystallize isomorphously. Anomalous scattering factors were deduced from synchrotron X-ray absorption measurements of crystals of the selenomethionyl proteins. Taken with reference to experience in the structural analysis of selenobiotinyl streptavidin by the method of multiwavelength anomalous diffraction (MAD), these data indicate that recombinant selenomethionyl proteins analyzed by MAD phasing offer a rather general means for the elucidation of atomic structures.

MeSH Terms
Chemical Phenomena Chemistry Crystallization Cystine/analogs & derivatives,metabolism Escherichia coli/metabolism Methionine/deficiency Models, Theoretical Organoselenium Compounds Protein Biosynthesis Proteins Recombinant Proteins/biosynthesis Selenium/metabolism Selenomethionine/metabolism Selenoproteins Thioredoxins/biosynthesis X-Ray Diffraction/methods
Chemicals
Organoselenium Compounds Proteins Recombinant Proteins Selenoproteins selenocystine Cystine Thioredoxins Selenomethionine Methionine Selenium
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Hendrickson W A
Howard Hughes Medical Institute, Columbia University, New York, NY 10032.
Horton J R
LeMaster D M
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1990-05-00
Pages
1665-72
Language
English
Region
England
NLM ID
8208664
PMCID
PMC551863
Subset
IM
Grants
NIGMS NIH HHS · GM34102 · United States
External Links
PubMed source PMC Full Text View journal details
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