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

2021 ◽  
Author(s):  
Ali Imran ◽  
Brandon S. Moyer ◽  
Ashley J. Canning ◽  
Dan Kalina ◽  
Thomas M Duncan ◽  
...  
Keyword(s):  
Binding Site ◽  
Primary Role ◽  
Dissociation Kinetics ◽  
High Affinity ◽  
Histone 3 ◽  
Quantitative Understanding ◽  
Slow Dissociation ◽  
Kinetics Of ◽  
Association Kinetics

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.

Characterization of a Specific, High Affinity [3H]Arginine8Vasopressin-Binding Site on Liver Microsomes from Different Strains of Rat and the Role of Magnesium*

Endocrinology ◽  
1986 ◽  
Vol 118 (3) ◽  
pp. 990-998 ◽  
Author(s):  
VENKAT GOPALAKRISHNAN ◽  
CHRIS R. TRIGGLE ◽  
PRAKASH V. SULAKHE ◽  
J. ROBERT McNEILL
Keyword(s):  
Binding Site ◽  
Liver Microsomes ◽  
High Affinity ◽  
Different Strains

Role of the mobility of antigen binding site in high affinity antibody elucidated by surface plasmon resonance

2016 ◽  
Vol 161 (1) ◽  
pp. 37-43 ◽  
Author(s):  
Natsuki Fukuda ◽  
Yoshiaki Suwa ◽  
Makiyo Uchida ◽  
Yoshihiro Kobashigawa ◽  
Hideshi Yokoyama ◽  
...  
Keyword(s):  
Surface Plasmon Resonance ◽  
Binding Site ◽  
Surface Plasmon ◽  
Plasmon Resonance ◽  
Antigen Binding ◽  
Antigen Binding Site ◽  
High Affinity

scholarly journals Pivotal role of α2 Na+pumps and their high affinity ouabain binding site in cardiovascular health and disease

2016 ◽  
Vol 594 (21) ◽  
pp. 6079-6103 ◽  
Author(s):  
Mordecai P. Blaustein ◽  
Ling Chen ◽  
John M. Hamlyn ◽  
Frans H. H. Leenen ◽  
Jerry B. Lingrel ◽  
...  
Keyword(s):  
Binding Site ◽  
Cardiovascular Health ◽  
Pivotal Role ◽  
Ouabain Binding ◽  
High Affinity ◽  
Health And Disease

Role of Binding Site Loops in Controlling Nitric Oxide Release:  Structure and Kinetics of Mutant Forms of Nitrophorin 4†,‡

Biochemistry ◽  
2004 ◽  
Vol 43 (21) ◽  
pp. 6679-6690 ◽  
Author(s):  
Estelle M. Maes ◽  
Andrzej Weichsel ◽  
John F. Andersen ◽  
Donald Shepley ◽  
William R. Montfort
Keyword(s):  
Nitric Oxide ◽  
Binding Site ◽  
Nitric Oxide Release ◽  
Nitrophorin 4 ◽  
Mutant Forms ◽  
Kinetics Of

Studies on the interaction of glutamate dehydrogenase with aporheine

1984 ◽  
Vol 49 (9) ◽  
pp. 2001-2011
Author(s):  
Jan Kovář ◽  
Luděk Matyska
Keyword(s):  
Binding Site ◽  
Glutamate Dehydrogenase ◽  
Inhibitory Action ◽  
Positive Charge ◽  
Alkaline Media ◽  
Acidic Ph ◽  
High Affinity ◽  
Good Substrate ◽  
Kinetics Of

The effect was examined of two inhibitors, aporheine and thyroxine, on the kinetics of action of glutamate dehydrogenase in the presence of the coenzyme, NADH, in an inhibiting excess. from the results obtained it appears probable that these two competing ligands do not bind to the regulatory binding site of the enzyme. If the good substrate, glutamate, is replaced by a poor one, such as alanine, aporheine can behave under certain conditions as an activator; this phenomenon is most likely due to fact, like in other cases, that the association of alanine wit the enzyme-coenzyme complex is the slowest step of the reaction. The inhibitory action of aporheine in alkaline media is more complicated than its action at neutral and acidic pH. The sigmoid dependence of inhibition on aporheine concentration is best interpreted as a result of the induction of the second binding site of the enzyme subunit for aporheine after the binding of the first molecule of this ligand. The results obtained indicate the presence of a positive charge localized in the neighborhood of the binding site for the first aporheine molecule and also the important role played by some group of the enzyme with a pK-value about 8 during the induction of the second binding site with a relative high affinity for this ligand.

scholarly journals Protease-activated receptor-4 uses dual prolines and an anionic retention motif for thrombin recognition and cleavage

2003 ◽  
Vol 376 (3) ◽  
pp. 733-740 ◽  
Author(s):  
Suzanne L. JACQUES ◽  
Athan KULIOPULOS
Keyword(s):  
Active Site ◽  
Human Platelets ◽  
Optimal Placement ◽  
High Affinity ◽  
Steady State Kinetic ◽  
Thrombin Activation ◽  
Protease Activated Receptor 1 ◽  
Anionic Cluster ◽  
Slow Dissociation ◽  
Kinetics Of

Thrombin activation of human platelets is mediated by the high-affinity PAR1 (protease-activated receptor-1) and the low-affinity PAR4 receptor. PAR1 and PAR4 exhibit markedly disparate kinetics of activation that likely reflect differences in the macromolecular association of thrombin with their respective N-terminal extracellular domains (exodomains). Here we examine the mechanism of initial thrombin binding and cleavage of the high- and low-affinity PAR exodomains using steady-state kinetic analyses. We showed that the PAR4 exodomain lacks the functional hirudin-like sequence found in PAR1 and does not bind exosite I to cause allosteric activation or inhibition of thrombin. Instead, PAR4 contains an anionic cluster, Asp57…Asp59…Glu62…Asp65 (DDED), in its exodomain, which slows the dissociation of PAR4 from the cationic thrombin. The analogous anionic residues in the PAR1 exodomain do not influence affinity for thrombin. Although PAR4 is cleaved more slowly than PAR1 on the cell surface, peptides containing the PAR4 P4-P1 active-site-interacting sequence, Pro45-Ala-Pro-Arg (PAPR), are efficiently cleaved due to the optimal placement of dual prolines at positions P4 and P2. In comparison, thrombin has low affinity and slow cleavage rates for peptides that have a P3 proline as occurs in human PAR3. Thus, to compensate for the lack of exosite I binding, PAR4 utilizes proline residues in its P4-P1 sequence to provide high-affinity interactions with the active site and an anionic cluster to slow dissociation from the cationic thrombin.

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