Sml1p is a dimer in solution: characterization of denaturation and renaturation of recombinant Sml1p

Biochemistry
Vibha GuptaChris G Dealwis

Abstract

Sml1p is a small 104-amino acid protein from Saccharomyces cerevisiae that binds to the large subunit (Rnr1p) of the ribonucleotide reductase complex (RNR) and inhibits its activity. During DNA damage, S phase, or both, RNR activity must be tightly regulated, since failure to control the cellular level of dNTP pools may lead to genetic abnormalities, such as genome rearrangements, or even cell death. Structural characterization of Sml1p is an important step in understanding the regulation of RNR. Until now the oligomeric state of Sml1p was unknown. Mass spectrometric analysis of wild-type Sml1p revealed an intermolecular disulfide bond involving the cysteine residue at position 14 of the primary sequence. To determine whether disulfide bonding is essential for Sml1p oligomerization, we mutated the Cys14 to serine. Sedimentation equilibrium measurements in the analytical ultracentrifuge show that both wild-type and C14S Sml1p exist as dimers in solution, indicating that the dimerization is not a result of a disulfide bond. Further studies of several truncated Sml1p mutants revealed that the N-terminal 8-20 residues are responsible for dimerization. Unfolding/refolding studies of wild-type and C14S Sml1p reveal that both proteins...Continue Reading

References

Jul 1, 1989·European Journal of Biochemistry·J A KnottA Weston
Sep 1, 1995·Trends in Biochemical Sciences·R A Sayle, E J Milner-White
Jul 4, 1997·The Journal of Biological Chemistry·A K MachadoG F Merrill
Oct 6, 1998·Annual Review of Biochemistry·A Jordan, P Reichard
Dec 14, 1999·The Journal of Biological Chemistry·A ChabesL Thelander
Aug 5, 2000·Journal of Molecular Biology·C BystroffD Baker
Oct 29, 2003·Biophysical Journal·Paulo Roberto LouzadaSérgio T Ferreira
Dec 10, 2003·The Journal of Biological Chemistry·Frederic RousseauLaura S Itzhaki
Dec 20, 2003·The Journal of Biological Chemistry·Tomoaki UchikiChris Dealwis

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Citations

Apr 21, 2005·Analytical Biochemistry·Joshua S SharpRobert L Hettich
Dec 17, 2008·Biochemistry·Jens DanielssonBirthe B Kragelund
May 13, 2006·Annual Review of Biophysics and Biomolecular Structure·Keiji Takamoto, Mark R Chance
Mar 16, 2006·Proceedings of the National Academy of Sciences of the United States of America·Hai XuChris Dealwis
Apr 8, 2014·Seminars in Cell & Developmental Biology·Estrella GuarinoStephen E Kearsey
Feb 25, 2020·Chemosphere·Olga Marchut-MikolajczykKatarzyna Struszczyk-Świta

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