Analysis of binding interfaces of the human scaffold protein AXIN1 by peptide microarrays

The Journal of Biological Chemistry
Jakub HarnošVítězslav Bryja

Abstract

Intrinsically disordered regions (IDRs) are protein regions that lack persistent secondary or tertiary structure under native conditions. IDRs represent >40% of the eukaryotic proteome and play a crucial role in protein-protein interactions. The classical approach for identification of these interaction interfaces is based on mutagenesis combined with biochemical techniques such as coimmunoprecipitation or yeast two-hybrid screening. This approach either provides information of low resolution (large deletions) or very laboriously tries to precisely define the binding epitope via single amino acid substitutions. Here, we report the use of a peptide microarray based on the human scaffold protein AXIN1 for high-throughput and -resolution mapping of binding sites for several AXIN1 interaction partners in vitro For each of the AXIN1-binding partners tested, i.e. casein kinase 1 ϵ (CK1ϵ); c-Myc; peptidyl-prolyl cis/trans isomerase, NIMA-interacting 1 (Pin1); and p53, we found at least three different epitopes, predominantly in the central IDR of AXIN1. We functionally validated the specific AXIN1-CK1ϵ interaction identified here with epitope-mimicking peptides and with AXIN1 variants having deletions of short binding epitopes. On the...Continue Reading

References

May 16, 2000·The EMBO Journal·K E SpinkW I Weis
Jan 4, 2001·Nature Genetics·R J ChoD J Lockhart
Jun 27, 2001·The Journal of Biological Chemistry·M KishidaA Kikuchi
Jul 5, 2001·Developmental Biology·R M McKayJ M Graff
Mar 9, 2002·The Journal of Biological Chemistry·Yi ZhangSheng-Cai Lin
Mar 30, 2002·Science·Rachel B BremLeonid Kruglyak
Jul 26, 2002·The Biochemical Journal·Alexsandra EspejoMark T Bedford
Feb 20, 2003·Protein Science : a Publication of the Protein Society·Livia OtteHartmut Oschkinat
Jan 23, 2004·PLoS Biology·Christiane LandgrafGianni Cesareni
Feb 18, 2004·Molecular and Cellular Biology·Feng CongHarold Varmus
Apr 7, 2004·Neuro-Signals·Wen Luo, Sheng-Cai Lin
Jun 2, 2004·Cellular and Molecular Life Sciences : CMLS·V BryjaA Hampl
Apr 17, 2007·Protein Expression and Purification·Jens ReinhardtLaurent Meijer
Jul 16, 2008·Progress in Biophysics and Molecular Biology·Marc S CorteseA Keith Dunker
Jan 10, 2009·The EMBO Journal·Hugh K ArnoldRosalie C Sears
Jan 27, 2010·Biochimica Et Biophysica Acta·Bin XueVladimir N Uversky
Nov 23, 2010·Journal of Molecular Biology·Maria NoutsouMadelon M Maurice
Jan 20, 2011·Proceedings of the National Academy of Sciences of the United States of America·Marc FiedlerMariann Bienz
Feb 3, 2011·The Journal of Biological Chemistry·Ondrej BernatikVitezslav Bryja
May 10, 2011·Science·Matthew C GoodWendell A Lim
Jul 6, 2014·The Journal of Biological Chemistry·Ondřej BernatíkVítězslav Bryja
Dec 23, 2014·Nature Reviews. Molecular Cell Biology·Peter E Wright, H Jane Dyson
Apr 1, 2013·Intrinsically Disordered Proteins·A Keith DunkerVladimir N Uversky

❮ Previous
Next ❯

Citations

Feb 21, 2019·Proceedings of the National Academy of Sciences of the United States of America·Michaela Kunova BosakovaPavel Krejci
Dec 12, 2020·The Biochemical Journal·Luke J Fulcher, Gopal P Sapkota
Jul 13, 2021·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Renata Erbert ContricianiLúcia Elvira Alvares

❮ Previous
Next ❯

Related Concepts

Related Feeds

Adherens Junctions

An adherens junction is defined as a cell junction whose cytoplasmic face is linked to the actin cytoskeleton. They can appear as bands encircling the cell (zonula adherens) or as spots of attachment to the extracellular matrix (adhesion plaques). Adherens junctions uniquely disassemble in uterine epithelial cells to allow the blastocyst to penetrate between epithelial cells. Discover the latest research on adherens junctions here.

Cadherins and Catenins

Cadherins (named for "calcium-dependent adhesion") are a type of cell adhesion molecule (CAM) that is important in the formation of adherens junctions to bind cells with each other. Catenins are a family of proteins found in complexes with cadherin cell adhesion molecules of animal cells: alpha-catenin can bind to β-catenin and can also bind actin. β-catenin binds the cytoplasmic domain of some cadherins. Discover the latest research on cadherins and catenins here.

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.