PMID: 9177186Jun 10, 1997Paper

Engineering subunit association of multisubunit proteins: a dimeric streptavidin

Proceedings of the National Academy of Sciences of the United States of America
T SanoC R Cantor

Abstract

A dimeric streptavidin has been designed by molecular modeling using effective binding free energy calculations that decompose the binding free energy into electrostatic, desolvation, and side chain entropy loss terms. A histidine-127 --> aspartic acid (H127D) mutation was sufficient to introduce electrostatic repulsion between subunits that prevents the formation of the natural tetramer. However, the high hydrophobicity of the dimer-dimer interface, which would be exposed to solvent in a dimeric streptavidin, suggests that the resulting molecule would have very low solubility in aqueous media. In agreement with the calculations, a streptavidin containing the H127D mutation formed insoluble aggregates. Thus, the major design goal was to reduce the hydrophobicity of the dimer-dimer interface while maintaining the fundamental structure. Free energy calculations suggested that the hydrophobicity of the dimer-dimer interface could be reduced significantly by deleting a loop from G113 through W120 that should have no apparent contact with biotin in a dimeric molecule. The resulting protein, containing both the H127D mutation and the loop deletion, formed a soluble dimeric streptavidin in the presence of biotin.

References

Aug 1, 1992·Trends in Biochemical Sciences·A Fersht, G Winter
Jan 1, 1992·Critical Reviews in Biotechnology·J S Sandhu
Apr 1, 1992·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·E S Ward
Jan 24, 1991·Nature·G Winter, C Milstein
Apr 30, 1991·Biochemical and Biophysical Research Communications·T Sano, C R Cantor
Jan 1, 1990·Methods in Enzymology·F W StudierJ W Dubendorff
Jan 1, 1990·Methods in Enzymology·N M Green
Jan 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·T Sano, C R Cantor
Jan 1, 1989·Advances in Immunology·S L Morrison, V T Oi
Apr 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·W A HendricksonR P Phizackerley
May 5, 1986·Journal of Molecular Biology·F W Studier, B A Moffatt
Jan 16, 1986·Nature·D Eisenberg, A D McLachlan
Feb 25, 1986·Nucleic Acids Research·C E ArgarañaC R Cantor
Apr 11, 1995·Proceedings of the National Academy of Sciences of the United States of America·T Sano, C R Cantor
Jan 1, 1995·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·R Raag, M Whitlow
Feb 28, 1995·Proceedings of the National Academy of Sciences of the United States of America·A ChilkotiP S Stayton
Feb 15, 1994·Proceedings of the National Academy of Sciences of the United States of America·T V BorchertR K Wierenga
Aug 1, 1995·Protein Engineering·L Demetrius
May 28, 1996·Proceedings of the National Academy of Sciences of the United States of America·P T Beernink, D R Tolan
Apr 1, 1996·Protein Science : a Publication of the Protein Society·Z WengC Delisi
Aug 1, 1996·Nature Biotechnology·G O ReznikT Sano
Dec 1, 1963·The Biochemical Journal·N M GREEN
Jul 20, 1964·Archives of Biochemistry and Biophysics·L CHAIET, F J WOLF

❮ Previous
Next ❯

Citations

Sep 1, 2005·Biotechnology Letters·Xipeng Liu, Jianhua Liu
Oct 29, 1998·Journal of Chromatography. B, Biomedical Sciences and Applications·T SanoC R Cantor
Apr 12, 2012·The Journal of Biological Chemistry·Amit MeirOded Livnah
Sep 21, 2012·Acta Crystallographica. Section D, Biological Crystallography·Pankaj PanwarEva Pebay-Peyroula
Apr 13, 2010·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Amit Meir, Oded Livnah
May 10, 2001·Protein Science : a Publication of the Protein Society·D L MeyerS S Graves
Nov 10, 2013·PloS One·Barbara TaskinenVesa P Hytönen
Jun 9, 2005·Proceedings of the National Academy of Sciences of the United States of America·Filiz M AslanCharles R Cantor
Nov 3, 2010·Journal of Molecular Graphics & Modelling·Cheng-Kuo Hsu, Sheldon Park
Sep 24, 2013·Journal of Molecular Biology·Michael FairheadMark Howarth
Sep 22, 2005·Proteins·Vesa P HytönenMarkku S Kulomaa
Jul 19, 2012·Biotechnology and Bioengineering·Kok Hong LimSheldon Park
Apr 20, 2011·Journal of Molecular Recognition : JMR·Jean-Marie TeulonJean-Luc Pellequer
Sep 23, 2008·Chemistry & Biology·Matthew Levy, Andrew D Ellington
Aug 10, 2004·Biophysical Journal·Mark J WanerDavid P Mascotti
May 15, 2007·Biophysical Journal·Denis B Tikhonov, Boris S Zhorov
Mar 22, 2001·Biophysical Journal·T CoussaertA P Gast
Nov 20, 2016·Photochemistry and Photobiology·Daria V SmirnovaNatalia N Ugarova
Nov 30, 2002·The Journal of Biological Chemistry·Olli H LaitinenMarkku S Kulomaa
Apr 29, 2020·American Journal of Transplantation : Official Journal of the American Society of Transplantation and the American Society of Transplant Surgeons·Pradeep ShresthaHaval Shirwan
Dec 21, 2002·The Journal of Biological Chemistry·Yael PazyOded Livnah
May 8, 2004·The Journal of Biological Chemistry·Henri R NordlundMarkku S Kulomaa
Aug 2, 2014·Bioconjugate Chemistry·Xun SunHaw Yang
Sep 10, 2021·Journal of the American Chemical Society·Bintian ZhangStuart Lindsay

❮ 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.