Kinetics of the multitasking high-affinity Win binding site of WDR5 in restricted and unrestricted conditions

Ali Imran; Brandon S Moyer; Ashley J Canning; Dan Kalina; Thomas M Duncan; Kelsey J Moody; Aaron J Wolfe; Michael S Cosgrove; Liviu Movileanu

doi:10.1042/BCJ20210253

Kinetics of the multitasking high-affinity Win binding site of WDR5 in restricted and unrestricted conditions

Biochem J. 2021 Jun 11;478(11):2145-2161. doi: 10.1042/BCJ20210253.

Authors

Ali Imran¹, Brandon S Moyer², Ashley J Canning³, Dan Kalina^{2

4}, Thomas M Duncan³, Kelsey J Moody^{1

2

4}, Aaron J Wolfe^{1

2

4}, Michael S Cosgrove³, Liviu Movileanu^{1

5

6}

Affiliations

¹ Department of Physics, Syracuse University, 201 Physics Building, Syracuse, New York 13244-1130, U.S.A.
² Ichor Therapeutics, Inc., 2521 US Route 11, LaFayette, New York 13084, U.S.A.
³ Department of Biochemistry and Molecular Biology, State University of New York, Upstate Medical University, 4249 Weiskotten Hall, 766 Irving Avenue, Syracuse, New York 13210, U.S.A.
⁴ Department of Chemistry, State University of New York, College of Environmental Science and Forestry, 1 Forestry Dr., Syracuse, New York 13210, U.S.A.
⁵ The BioInspired Institute, Syracuse University, Syracuse, New York 13244, U.S.A.
⁶ Department of Biomedical and Chemical Engineering, Syracuse University, 329 Link Hall, Syracuse, New York 13244, U.S.A.

Abstract

Recent advances in quantitative proteomics show that WD40 proteins play a pivotal role in numerous cellular networks. Yet, they have been fairly unexplored and their physical associations with other proteins are ambiguous. A quantitative understanding of these interactions has wide-ranging significance. WD40 repeat protein 5 (WDR5) interacts with all members of human SET1/MLL methyltransferases, which regulate methylation of the histone 3 lysine 4 (H3K4). Here, using real-time binding measurements in a high-throughput setting, we identified the kinetic fingerprint of transient associations between WDR5 and 14-residue WDR5 interaction (Win) motif peptides of each SET1 protein (SET1Win). Our results reveal that the high-affinity WDR5-SET1Win interactions feature slow association kinetics. This finding is likely due to the requirement of SET1Win to insert into the narrow WDR5 cavity, also named the Win binding site. Furthermore, our explorations indicate fairly slow dissociation kinetics. This conclusion is in accordance with the primary role of WDR5 in maintaining the functional integrity of a large multisubunit complex, which regulates the histone methylation. Because the Win binding site is considered a key therapeutic target, the immediate outcomes of this study could form the basis for accelerated developments in medical biotechnology.

Keywords: SET1/MLL proteins; biolaye interferometry; fluorescence polarization; mixed lineage leukemia; protein–peptide interactions; surface plasmon resonance.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Amino Acid Motifs
Binding Sites
Histone-Lysine N-Methyltransferase / chemistry
Histone-Lysine N-Methyltransferase / genetics
Histone-Lysine N-Methyltransferase / metabolism*
Humans
Intracellular Signaling Peptides and Proteins / chemistry
Intracellular Signaling Peptides and Proteins / genetics
Intracellular Signaling Peptides and Proteins / metabolism*
Kinetics
Peptide Fragments / chemistry
Peptide Fragments / genetics
Peptide Fragments / metabolism*
Protein Binding
Protein Conformation
Protein Interaction Domains and Motifs*

Substances

Intracellular Signaling Peptides and Proteins
Peptide Fragments
WDR5 protein, human
Histone-Lysine N-Methyltransferase
Setd1A protein, human

Abstract

Publication types

MeSH terms

Substances

Grants and funding