ANKUSH GOYAL

JR 2

GMC PATIALA

Pharmacology of Enzyme-Linked Receptors

Enzyme-linked receptors, also known as receptor tyrosine kinases (RTKs) or

single-transmembrane-span catalytic receptors, are a class of cell-surface receptors that
mediate cellular responses via enzymatic activity on their cytoplasmic domain following
ligand binding.

I. Structure

●​ Single polypeptide chain with:

o​ Extracellular ligand-binding domain
o​ Transmembrane domain
o​ Intracellular catalytic domain (intrinsic or associated enzymatic activity)

II. Types of Enzyme-Linked Receptors

1.​ Receptor Tyrosine Kinases (RTKs)

o​ E.g.: Insulin receptor, EGFR (epidermal growth factor receptor)
2.​ Receptor Serine/Threonine Kinases
o​ E.g.: TGF-β receptor
3.​ Receptor Guanylyl Cyclases
o​ E.g.: ANP receptor
4.​ Receptors Associated with Tyrosine Kinases (Non-receptor tyrosine kinases)
o​ E.g.: Cytokine receptors like IL-2, erythropoietin receptor (via JAK/STAT
pathway)
5.​ Receptor Tyrosine Phosphatases
o​ Less understood, mainly in immune regulation

III. Mechanism of Action (MOA)

1.​ Ligand binding to extracellular domain

 2.​ Receptor dimerization (except in some cytokine receptors)
3.​ Autophosphorylation of tyrosine residues on intracellular domain
4.​ Recruitment of adaptor proteins (e.g., Grb2, SOS)
5.​ Activation of downstream signaling cascades:
o​ MAPK pathway (growth, proliferation)
o​ PI3K/Akt pathway (survival, metabolism)
o​ JAK/STAT pathway (transcriptional regulation)

IV. Pharmacological Modulation

1. Agonists

●​ Insulin: Activates insulin receptor (RTK)

●​ Atrial Natriuretic Peptide (ANP): Stimulates guanylyl cyclase receptors

2. Antagonists / Inhibitors

●​ Monoclonal antibodies:
o​ Trastuzumab: HER2/neu receptor antagonist
o​ Cetuximab: EGFR inhibitor
●​ Tyrosine kinase inhibitors (TKIs):
o​ Imatinib: BCR-ABL and c-Kit inhibitor
o​ Erlotinib, Gefitinib: EGFR TKIs

V. Clinical Significance

●​ Cancer: Dysregulated RTK signaling in breast, lung, CML

●​ Diabetes: Impaired insulin receptor signaling
●​ Cardiovascular: ANP receptor activation used in heart failure
●​ Immunology: JAK inhibitors (e.g., Tofacitinib) in autoimmune disorders

VI. Diagram (if allowed in your exam – always draw to get bonus marks)

Include a labeled schematic showing:

●​ Ligand binding
●​ Dimerization
●​ Autophosphorylation
●​ Downstream signaling pathways (MAPK, PI3K)
VII. Recent Advances

●​ Biologics: E.g., monoclonal antibodies targeting specific RTKs

●​ Precision medicine: TKIs based on mutational profiling (e.g., EGFR mutations in
NSCLC)
●​ Gene therapy approaches for receptor modification

Conclusion

Enzyme-linked receptors play a pivotal role in regulating cell growth, differentiation,

metabolism, and immune responses. Their pharmacological modulation has revolutionized
treatment paradigms, especially in oncology and metabolic diseases

Kinase-Linked and Related Receptors

🧬 General Features
●​ Structure: Large single-chain transmembrane proteins (~1000 amino acids)
o​ One extracellular ligand-binding domain
o​ One transmembrane helical segment
o​ One intracellular domain with enzymatic activity or docking sites
●​ Mechanism: Ligand binding → receptor dimerization → activation of intracellular
kinase domain → phosphorylation of target proteins → gene transcription changes
●​ Ligands: Protein mediators (growth factors, cytokines), hormones (e.g. insulin, leptin)
●​ Functions: Regulate gene transcription, cell growth, division, differentiation,
metabolism, immunity, apoptosis, and tissue repair

🔬 Main Classes of Kinase-Linked Receptors

1️⃣ Receptor Tyrosine Kinases (RTKs)

●​ Key Feature: Intrinsic tyrosine kinase activity in intracellular domain

●​ Examples:
o​ Epidermal Growth Factor Receptor (EGFR)
o​ Nerve Growth Factor Receptor (NGFR)
o​ Insulin Receptor (dimeric; indirectly activates TK)
o​ Toll-like Receptors (TLRs) (recognize bacterial LPS; role in innate immunity)
 ●​ Mechanism:
o​ Ligand binding → dimerization → autophosphorylation on tyrosine residues →
downstream signaling (e.g., MAPK, PI3K-Akt)

2️⃣ Receptor Serine/Threonine Kinases

●​ Key Feature: Intrinsic kinase activity that phosphorylates serine or threonine residues
●​ Main Example: Transforming Growth Factor-β (TGF-β) receptor
●​ Function: Cell proliferation, differentiation, and immune modulation

3️⃣ Cytokine Receptors

●​ Key Feature: Lack intrinsic kinase activity; rely on associated kinases

●​ Key Kinase: Janus kinase (JAK)
●​ Ligands:
o​ Interferons
o​ Colony-Stimulating Factors (CSFs)
o​ Other cytokines
●​ Pathway: Ligand binding → JAK activation → STAT phosphorylation → gene
transcription

📚 Clinical Relevance
●​ Cancer therapy: Targeting RTKs (e.g., EGFR inhibitors in cancer)
●​ Autoimmune diseases: JAK inhibitors for rheumatoid arthritis
●​ Diabetes: Insulin signaling via RTK pathway
●​ Fibrosis & Cancer: TGF-β pathway as target

🔁 Comparison with Other Receptor Types

Feature Ligand-Gated Ion Channel GPCR Kinase-Linked Receptors
Signal Onset Very fast (ms) Fast (s) Slow (minutes to hours)
Coupling Kinases (intrinsic or
Ion channel G-proteins
Mechanism associated)
cAMP, Gene transcription, protein
Major Outcome Membrane potential change
IP3/DAG synthesis
 Feature Ligand-Gated Ion Channel GPCR Kinase-Linked Receptors
GABA_A,
Examples β-adrenergic Insulin, EGF, TGF, cytokines
NMDA

📌 Kinase-Linked Receptors (Enzyme-Linked Receptors)

🔷 Common Structural Features
●​ Single transmembrane protein:
o​ Large extracellular ligand-binding domain
o​ Single membrane-spanning helix
o​ Intracellular domain with enzymatic activity (or binds kinases)
●​ Ligand binding → Receptor dimerization → Autophosphorylation of tyrosine
residues

🔬 Mechanism of Action
1.​ Ligand Binding → Dimerization
2.​ Autophosphorylation on tyrosine residues (or activation of associated kinases like JAK)
3.​ Phosphotyrosines act as docking sites for SH2-domain proteins (Src Homology 2
domain)
4.​ These proteins activate downstream cascades:
o​ Kinase cascades
o​ Phospholipase activation
o​ Transcription factor activation

🧬 Functions
●​ Control of:
o​ Cell growth and division
o​ Differentiation
o​ Gene expression
o​ Inflammatory mediator release

📈 Key Signal Transduction Pathways

✅ 1. Ras/Raf/MAP Kinase Pathway
●​ Activated mainly by growth factors
●​ Sequence:
1.​ SH2-domain protein Grb2 binds phosphorylated receptor
2.​ Ras (a proto-oncogene, G-protein-like) is activated via GDP-GTP exchange
3.​ Ras → Raf → MEK → MAP kinase (serine/threonine kinases)
4.​ MAP kinase activates transcription factors → gene expression
●​ Involved in:

o​ Cell proliferation
o​ Differentiation
o​ Cancer, inflammation, neurodegeneration

✅ 2. Jak/Stat Pathway
●​ Activated mainly by cytokines
●​ Steps:
1.​ Cytokine binds → receptor dimerizes
2.​ Jak (Janus kinase, cytosolic TK) binds receptor → phosphorylates it
3.​ Stat (Signal Transducer and Activator of Transcription) binds via SH2 domain
4.​ Stat is phosphorylated → dimerizes → enters nucleus → activates gene
transcription
●​ Key in inflammation and immune responses

🔁 Other Relevant Pathways

✅ PI3-Kinase Pathway
●​ Activated by RTKs and GPCRs
●​ Converts PIP2 → PIP3
●​ PIP3 acts as a docking lipid for proteins like Akt/PKB
●​ Role: Cell survival, metabolism, and growth

🧬 SH2 Domain Proteins

●​ ~100 amino acid motif
●​ Bind selectively to phosphotyrosine residues
●​ Examples: kinases, phospholipases, adaptor proteins
 ●​ Specificity gives unique signaling patterns for different ligands

🔥 Clinical and Research Notes

●​ Src oncogene (v-Src) from Rous sarcoma virus encodes a tyrosine kinase; related to host
c-Src
o​ First oncogene discovered (1979)
●​ NF-κB Pathway:
o​ Inactive in cytosol (bound to IκB)
o​ IKK activation → IκB phosphorylation → NF-κB released → enters nucleus
→ activates inflammatory genes
●​ Protein kinase B (Akt/PKB): Named after PKA and PKC; key in PI3K pathway

🧪 Pharmacological Importance
●​ RTKs targeted in cancer therapy (e.g., EGFR inhibitors)
●​ Jak inhibitors used in autoimmune diseases (e.g., rheumatoid arthritis)
●​ PI3K/Akt pathway is a target in oncology and metabolic disorders
●​ Mutations in Ras/Raf/MAPK cascade → cancer, often due to constitutive activation

Here are concise, exam-oriented notes on Protein Kinase Cascades and Phosphorylation
Mechanisms from the given text, tailored for MD Pharmacology:

📌 Protein Phosphorylation in Signal Transduction

🔬 Core Concepts
●​ Protein phosphorylation is a universal mechanism for regulating:
o​ Enzymes
o​ Ion channels
o​ Transporters
o​ Receptors
o​ Transcription factors
●​ It is reversible, controlled by:
o​ Kinases (add phosphate)
o​ Phosphatases (remove phosphate)
🔗 Kinase Cascades ("Signal Amplifiers")
●​ Ligand binding to receptors activates kinase cascades, which lead to amplification and
diversity in cellular responses.
●​ Receptor types activating kinases:
o​ GPCRs
o​ Kinase-linked receptors (e.g., RTKs)
o​ Guanylyl cyclase-linked receptors (via cGMP)

📈 Mechanism of Amplification
1.​ Receptor activation (e.g., by hormone or cytokine)
2.​ Activation of first kinase in the cascade
3.​ Sequential phosphorylation of downstream kinases (e.g., Raf → MEK → MAPK)
4.​ Regulation of multiple intracellular target proteins:
o​ Transcription factors
o​ Enzymes
o​ Cytoskeletal proteins
o​ Channels
o​ Secretory mechanisms

🧬 Key Players in Signal Cascades

●​ Protein Kinase A (PKA) – cAMP-dependent
●​ Protein Kinase C (PKC) – DAG & Ca²⁺-dependent
●​ Protein Kinase G (PKG) – cGMP-dependent
●​ Protein Kinase B (PKB/Akt) – activated via PIP3, controls:
o​ Apoptosis inhibition
o​ Proliferation
o​ Differentiation
o​ Endothelial NO synthase (eNOS) activation

🧪 Specificity in Signaling
●​ Over 500 different kinases in the human genome
●​ Each kinase has distinct substrate specificity​
→ ensures selective activation of downstream responses for each receptor/hormone
🧘 Feedback & Cross-Talk
●​ Most kinases and receptors are themselves regulated by phosphorylation
●​ Cross-talk between pathways enables integration of signals
●​ Desensitization:
o​ GPCR kinases (GRKs) phosphorylate GPCRs → internalization and reduced
responsiveness

🔄 Deactivation Mechanism
●​ Large family of protein phosphatases:
o​ Reverses kinase action
o​ Restores basal activity
o​ Controls signal duration and intensity

💊 Clinical Relevance
●​ Imatinib: First designed kinase inhibitor, targets BCR-Abl tyrosine kinase in chronic
myeloid leukemia (CML)
●​ Protein kinases are major drug targets in:
o​ Cancer
o​ Inflammation
o​ Autoimmune diseases

🧠 Concept Shift in Pharmacology

●​ The older "receptor → effect" paradigm is being replaced by:
o​ Complex networks of intracellular signaling
o​ Systems pharmacology approach to understand drug action