Design of a leucine zipper coiled coil stabilized 1.4 kcal mol-1 by phosphorylation of a serine in the e position

Protein Science : a Publication of the Protein Society
L SzilákC Vinson

Abstract

Using a dimeric bZIP protein, we have designed a leucine zipper that becomes more stable after a serine in the e position is phosphorylated by protein kinase A (delta delta GP = -1.4 kcal mol-1 dimer-1 or -0.7 kcal mol-1 residue-1). Mutagenesis studies indicate that three arginines form a network of inter-helical (i,i' + 5; i, i' + 2) and intra-helical (i, i + 4) attractive interactions with the phosphorylated serine. When the arginines are replaced with lysines, the stabilizing effect of serine phosphorylation is reduced (delta delta GP = -0.5 kcal mol-1 dimer-1). The hydrophobic interface of the leucine zipper needs a glycine in the d position to obtain an increase in stability after phosphorylation. The phosphorylated protein binds DNA with a 15-fold higher affinity. Using a transient transfection assay, we document a PKA dependent four-fold activation of a reporter gene. Phosphorylation of a threonine in the same e position decreases the stability by delta delta GP = +1.2 kcal mol-1 dimer-1. We present circular dichroism (CD) thermal denaturations of 15 bZIP proteins before and after phosphorylation. These data provide insights into the structural determinants that result in stabilization of a coiled coil by phosphorylation.

References

Dec 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·F SangerA R Coulson
Oct 25, 1975·Journal of Molecular Biology·A D McLachlan, M Stewart
Dec 20, 1990·Journal of Molecular Biology·A HorovitzA R Fersht
Dec 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·S Marqusee, R L Baldwin
May 5, 1986·Journal of Molecular Biology·F W Studier, B A Moffatt
Jan 1, 1984·Annual Review of Biophysics and Bioengineering·A A Zamyatnin
Apr 1, 1993·Current Opinion in Genetics & Development·A D Baxevanis, C R Vinson
Jun 1, 1996·Nature Structural Biology·L GonzalezT Alber
Feb 1, 1997·Nature Structural Biology·L SzilákC Vinson

❮ Previous
Next ❯

Citations

Nov 5, 2003·Journal of Molecular Biology·Rachel S Signarvic, William F DeGrado
Feb 13, 2001·Trends in Cell Biology·P BurkhardS V Strelkov
Sep 12, 2009·Journal of the American Chemical Society·Alexander J Riemen, Marcey L Waters
Feb 13, 2003·Proceedings of the National Academy of Sciences of the United States of America·A Soren LeonardMichael J Welsh
Nov 23, 2000·Proceedings of the National Academy of Sciences of the United States of America·J R NewmanP S Kim
May 25, 2002·Molecular and Cellular Biology·Xuewen Pan, Joseph Heitman
Oct 13, 2007·Molecular Endocrinology·Bin HeBert W O'Malley
May 23, 2012·Biochimica Et Biophysica Acta·Yamini KrishnanThomas V McDonald
Feb 5, 2014·Journal of Nanobiotechnology·Helena Gradišar, Roman Jerala
Nov 20, 2012·Structure·Colin A SmithTanja Kortemme
Mar 3, 2007·Faraday Discussions·Israel Fernández, Gernot Frenking
Aug 16, 2002·Molecular Microbiology·Robin M Delahay, Gad Frankel
May 18, 2010·Angewandte Chemie·Hana Robson Marsden, Alexander Kros
May 21, 2010·Organic & Biomolecular Chemistry·Malgorzata BroncelBeate Koksch
Sep 22, 2010·Organic & Biomolecular Chemistry·Alexander J Riemen, Marcey L Waters
Jul 26, 2014·Biomedical Papers of the Medical Faculty of the University Palacký, Olomouc, Czechoslovakia·Petr HabalJiri Mandak
Jun 14, 2002·American Journal of Physiology. Heart and Circulatory Physiology·Qianxun XiaoKaie Ojamaa
Sep 1, 2020·Angewandte Chemie·Valerio FasanoVarinder K Aggarwal
Jun 17, 2020·Current Research in Structural Biology·Zhou YinGabby Rudenko
Mar 7, 2021·Nature Communications·Leon HarringtonPetra Schwille
Nov 30, 2019·Journal of Structural Biology·Komal ChoukateBarnali Chaudhuri
Apr 7, 2010·Journal of the American Chemical Society·Susan Carr ZondloNeal J Zondlo
Jun 12, 2010·Journal of the American Chemical Society·Alexander J Riemen, Marcey L Waters
Mar 25, 2011·Analytical Chemistry·Lucrèce MatheronBolbach Gèrard
Jul 24, 2020·ACS Synthetic Biology·Daniel L WinterDominic J Glover
Aug 24, 2021·ACS Pharmacology & Translational Science·Omid Tavassoly, Iman Tavassoly
Apr 28, 2006·Journal of the American Chemical Society·Shalini Balakrishnan, Neal J Zondlo

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.