SEMINAR PRESENTATION

ON

ALLOSTERIC MECHANISM OF SIGNAL

TRANSDUCTION
BY

TINUOLA MARIAM OLANIYI

(20/57BC/01167)
DEPARTMENT OF MEDICAL BIOCHEMISTRY AND PHARMACOLOGY,
FACULTY OF PURE AND APPLIED SCIENCES.
KWARA STATE UNIVERSITY, MALETE, NIGERIA.
1
OUTLINE
 INTRODUCTION
 ALLOSTERY
 SIGNIFICANCE OF ALLOSTERIC REGULATION IN SIGNAL TRANSDUCTION
 ALLOSTERIC MODULATORS
 ALLOSTERIC SITE
 FEATURES OF ALLOSTERIC ENZYMES
 ALLOSTERIC ACTIVATION
 ALLOSTERIC INHIBITION
 G PROTEIN-COUPLED RECEPTORS (GPCRS)
 LIGAND-GATED ION CHANNELS
 SIGNAL TRANSDUCTION PATHWAYS INVOLVING ALLOSTERIC RECEPTORS
 ALLOSTERIC REGULATION IN CANCER
 CONCLUSION
 REFERENCES
 INTRODUCTION

 Proteins, essential macromolecules, have complex structures with multiple ligand-binding sites. Enzymes

showcase intricate architecture with active and allosteric sites playing pivotal roles (Csermely et al., 2017).

 Active site: Central to enzymatic activity, binds substrates, and dictates protein functionality (Deribe et al., 2019).

 Allosteric site: Serves as a docking point for allosteric effectors, influencing reaction rates and binding affinity

(Hilser et al., 2017).

 Allosteric effector binding induces positive/negative influences, triggering a conformational change (Kenakin,

2017).

 Allosteric studies explore dynamic aspects, including conformational transitions and alterations in substrate

binding affinity (Kenakin, 2017).

 ALLOSTERY

 The term "allostery," stemming from the Greek "allo," meaning "other," and "stereos," meaning "solid" or
"object," is a fundamental concept in biochemistry that intricately governs protein regulation within living
cells (Joseph et al., 2017).

 Allosteric proteins, exemplified by notable instances like hemoglobin, showcase a remarkable

adaptability, responding intelligently to fluctuations in environmental stimuli, particularly changes in
ligand concentrations such as ions, metabolites, and macromolecules (Joseph et al., 2017).

 The essence of allostery lies in its ability to orchestrate events in one part of a molecule, instigating
consequential effects in another region (Kuriyan and Eisenberg, 2018).
SIGNIFICANCE OF ALLOSTERIC REGULATION IN SIGNAL
TRANSDUCTION

 INTEGRATION OF SIGNALS

 AMPLIFICATION OF SIGNALS

 FINE-TUNING CELLULAR RESPONSES

 DYSREGULATION AND DISEASE: (Boehr et al., 2016).

 ALLOSTERIC MODULATORS

 In biochemistry, allosteric modulators represent a category of compounds that engage with receptors,
inducing alterations in the receptors' responses to external stimuli (Popovych et al., 2018).
 Some notable examples of such modulators include benzodiazepines and alcoholic beverages, both of which
exhibit psychoactive properties (Popovych et al., 2018).
 Allosteric modulators manifest in three distinct types: positive, negative, and neutral (Lau et al., 2017).

 Positive Allosteric Modulators (PAMs)

 Negative Allosteric Modulators (NAMs)
 Neutral Allosteric Modulators
 ALLOSTERIC SITE

 The allosteric site represents a pivotal regulatory locus within an enzyme, providing a binding site for effector

molecules that are distinct from the enzyme's substrate (Lechtenberg et al., 2019).

 These effector molecules can either activate or inhibit the catalytic efficiency of the enzyme (Lechtenberg et al.,

2019).

 The dynamic interaction between effectors and the enzyme's allosteric site induces conformational changes in the

enzyme, thereby either facilitating or impeding substrate binding to the catalytic site (Lechtenberg et al., 2019).
 FEATURES OF ALLOSTERIC ENZYMES

 QUATERNARY STRUCTURE

 REACTION RATE DYNAMICS

 SPATIAL SEPARATION

 CONFORMATIONAL STATES

 MULTIPLE BINDING SITES

 DUAL FUNCTIONALITY (Lechtenberg et al., 2019).

Figure 1: Showing the Allosteric site (RED outline).
Source: (Maksay, 2016).
 ALLOSTERIC ACTIVATION

 Allosteric activation occurs when an effector molecule binds to the allosteric site, inducing a conformational change
in the enzyme (Taylor et al., 2015).

 The binding of an allosteric activator, often a specific molecule, triggers enhanced affinity between the enzyme and
its substrate (Sinha and Nussinov, 2015).

 Allosteric activation typically promotes the catalytic activity of the enzyme, ensuring a rapid and efficient cellular
response (Sinha and Nussinov, 2015).

 A notable example involves isoleucine acting as an allosteric activator, stimulating an enzyme crucial in its own
synthesis, thereby facilitating the production of essential cellular components (Nussinov et al., 2018).
 ALLOSTERIC INHIBITION

 Allosteric inhibition involves the binding of molecules to allosteric sites, leading to conformational changes that

dampen protein activity (Zhuravlev and Papoian, 2019).

 Citrate inhibition of phosphofructokinase in glycolysis typifies this process, where citrate, an intermediate in the

citric acid cycle, acts as an allosteric inhibitor, regulating the pace of glycolytic flux to prevent excessive energy

production (Zhuravlev and Papoian, 2019).

 Allosteric regulation is integral to maintaining cellular homeostasis, providing a swift and reversible means for

organisms to adapt to fluctuating conditions (Zhuravlev and Papoian, 2019).

Figure 2: Illustration of Allosteric Inhibition and Allosteric Activation.
Source: (Nussinov et al., 2018).
 G Protein-Coupled Receptors (GPCRs)

 Constitute a substantial portion of allosteric receptors, playing a pivotal role in diverse physiological processes
(Tsai et al., 2016).
 Structurally, GPCRs are integral membrane proteins with seven transmembrane helices (Tsai et al., 2016).

 Common GPCR Signaling Pathways (Adrenergic Receptors):

 GPCRs, such as β-adrenergic receptors, respond to ligands like epinephrine. Activation leads to the
stimulation of adenylate cyclase through the Gαs subunit (Nussinov et al., 2018).
 Increased cyclic AMP (cAMP) production subsequently activates protein kinase A (PKA), initiating
diverse cellular responses (Tsai and Nussinov, 2018).
 Ligand-Gated Ion Channels

 hese receptors, often referred to as ionotropic receptors, directly control the flow of ions across the cell membrane
in response to ligand binding (Singh and Uversky, 2021).
 The activation of ligand-gated ion channels results in changes in membrane potential and ion concentrations,
crucial for neuronal signaling and other physiological processes (Xie and Vucetic, 2020).

 Common Ligand-Gated Ion Channels (Nicotinic Acetylcholine Receptors (nAChRs):

 nAChRs respond to acetylcholine, a neurotransmitter.

 Allosteric modulators, like positive allosteric modulators (PAMs), can enhance the response to
acetylcholine, influencing synaptic transmission (Stetz et al., 2020).
 SIGNAL TRANSDUCTION PATHWAYS INVOLVING ALLOSTERIC RECEPTORS
(ALLOSTERIC REGULATION IN CELL CYCLE CONTROL)

 Cell cycle control, orchestrating the division of cells, involves intricate regulatory mechanisms (Chatzigoulas and Cournia, 2022).

 Positive Regulation: Cyclins and CDKs

 Cyclins and Cdks form complexes to regulate the cell cycle, with cyclin levels fluctuating throughout phases (Wenthur et al., 2018).

 The phosphorylation of Cdk/cyclin complexes is crucial for their activation, allowing them to phosphorylate proteins involved in cell cycle

progression (Fang et al., 2017).

 The concentration of fully activated cyclin/Cdk complexes dictates cell cycle progression through checkpoints (Quaglia et al., 2023).

 Negative Regulation: Rb, p53, and p21

 Negative regulators, such as Rb, p53, and p21, act at the G1 checkpoint (Bah and Forman-Kay, 2018). p53, responsive to damaged DNA, halts

the cell cycle, promoting DNA repair or apoptosis (Jin and Grater, 2022).

 Rb monitors cell size, inhibiting transcription factors until phosphorylated (Nussinov et al., 2015).

 The coordinated action of positive and negative regulators ensures the orderly progression of the cell cycle, highlighting the intricate role of

allosteric regulation in cell division control (Stetz et al., 2020).

 ALLOSTERIC REGULATION IN CANCER

 GPCR kinases (GRKs) exemplify allosteric regulation in cancer, selectively recognizing distinct conformations of
GPCRs and initiating phosphorylation at specific residues. This process triggers downstream signaling events, such as
ERK 1/2 activation (Bah and Forman-Kay, 2018).

 The interplay between allosteric regulation and posttranslational modifications is critical for understanding the
nuanced signaling events driving oncogenesis. This complex interaction provides insights into the regulation of key
cellular processes, such as the activation of ERK 1/2 cascades (Jin and Grater, 2022).

 Allosteric mutations within kinase catalytic core domains can result in the constitutive activation of aberrant kinases,
contributing to oncogenesis. A comprehensive mechanistic understanding of these diverse allosteric mutations is
essential for the development of effective cancer therapies, highlighting the complexity of targeting allosteric sites in
drug design (Jin and Grater, 2022).
 CONCLUSION
 Allosteric modulators, including Positive Allosteric Modulators (PAMs), Negative Allosteric Modulators (NAMs), and Neutral Allosteric Modulators
(NAMs), act as conductors fine-tuning cellular activities. By binding to allosteric sites and inducing protein conformational changes, these molecules
contribute to precise control of cellular regulation.

 Examination of allosteric enzymes highlights their unique properties. Understanding how these enzymes respond to allosteric modulators, causing
conformational changes that either activate or inhibit catalytic activity, underscores their evolutionary significance in maintaining cellular
homeostasis.

 G-Protein Coupled Receptors (GPCRs), Ionotropic Receptors, and Tyrosine Kinase Receptors serve as molecular switches in cell signaling. Allosteric
regulation of these receptors influences signal transduction pathways, impacting diverse physiological processes.

 Allosteric regulation's broader implications span transcription factors, metabolism, cell cycle control, and neurotransmission. Positive and negative
regulations in contexts like cyclins, Cdks, Rb, p53, p21, NMDAR subunits, and associated modulators unveil the intricate tapestry of allosteric
regulation across various biological contexts.

 The seminar report emphasizes the impact of allosteric regulation in disease states, focusing on cancer and explores allosteric modulation as a
promising therapeutic avenue for targeted interventions and disease mitigation.
 References
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THANK
YOU