Rapid, optimized interactomic screening.
We must reliably map the interactomes of cellular macromolecular complexes in order to fully explore and understand biological systems. However, there are no methods to accurately predict how to capture a given macromolecular complex with its physiological binding partners. Here, we present a screening method that comprehensively explores the parameters affecting the stability of interactions in affinity-captured complexes, enabling the discovery of physiological binding partners in unparalleled detail. We have implemented this screen on several macromolecular complexes from a variety of organisms, revealing novel profiles for even well-studied proteins. Our approach is robust, economical and automatable, providing inroads to the rigorous, systematic dissection of cellular interactomes.
Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, New York, USA.
Zhanna
Z
Hakhverdyan
1] Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, New York, USA. [2] Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark.
Michal
M
Domanski
Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, New York, USA.
Loren E
LE
Hough
Orochem Technologies Inc., Naperville, Illinois, USA.
Asha A
AA
Oroskar
Orochem Technologies Inc., Naperville, Illinois, USA.
Anil R
AR
Oroskar
1] Center for Health Informatics and Bioinformatics, New York University School of Medicine, New York, New York, USA. [2] Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, New York, New York, USA.
Sarah
S
Keegan
1] Institute for Systems Biology, Seattle, Washington, USA. [2] Seattle Biomedical Research Institute, Seattle, Washington, USA.
David J
DJ
Dilworth
Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, New York, USA.
Kelly R
KR
Molloy
High Energy Physics Instrument Shop, The Rockefeller University, New York, New York, USA.
Vadim
V
Sherman
1] Institute for Systems Biology, Seattle, Washington, USA. [2] Seattle Biomedical Research Institute, Seattle, Washington, USA.
John D
JD
Aitchison
1] Center for Health Informatics and Bioinformatics, New York University School of Medicine, New York, New York, USA. [2] Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, New York, New York, USA.
David
D
Fenyö
Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, New York, USA.
Brian T
BT
Chait
Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark.
Torben Heick
TH
Jensen
Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, New York, USA.
Michael P
MP
Rout
Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, New York, USA.
John
0000000263077713
J
LaCava
D002460:Cell Line; D004926:Escherichia coli; D006801:Humans; D046911:Macromolecular Substances / Q000378:metabolism*; D025941:Protein Interaction Mapping / Q000379:methods*; D060066:Protein Interaction Maps; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D015003:Yeasts
D046911:Macromolecular Substances; D011506:Proteins