Signal Transduction

Dr. Chandi C Mandal, PhD

Phone No: 9650569396 Email ID: cc.mandal@sharda.ac.in

Syllabus of Signal Transduction

L T P: 3-1-2 Credit: 5 Unit I: G Protein-Coupled Signaling Pathways (10) Classification, structure and regulation of transmembrane receptors; G-protein-coupled receptors; Structure, ligand binding and signal transmission; Regulatory GTP-ases; Heterotrimeric G proteins; Second messengers: cAMP and cGMP. Unit II: Protein Kinases (10) Classification and characteristics of protein kinases; Protein phosphorylation and dephosphorylation; Regulation of Protein Kinase A pathway, Phosphatidyl-3-kinase (PI3K)/Akt pathway; Janus Kinase and Signal Transducer and Activator of Transcription (JAK-STAT) pathway; Protein cascades of MAPK; Organization of MAPK pathways. Unit III: Phosphatase and their Types (10) Role of phosphatase in signal transduction; Types of phosphatase: Tyrosine, serine/threonine, dual specificity phosphatase and their physiological relevance. Unit IV: Apoptosis (10) Classification and functions of caspases; Intrinsic and Extrinsic death pathways; Bcl-2 protein family; Role of p53 in apoptosis. Reference Books: Krauss G., Biochemistry of Signal Transduction and Regulation, Wiley-VCH, 2008. Hancock J.T., Cell Signalling, Oxford University Press, 2010. Gomperts B.D., Kramer I.M. and Tatham P.E.R., Signal Transduction, Academic Press, 2009.

General Function of Signal Pathways

The enormous structural varieties and functional capacities of multicellular organisms are due to their ability to coordinate the biochemical reactions of the various cells of the whole organisms. The basis for this coordination is the intercellular communication, which allows a single cell to influence the behavior and functional property of other cells in a specific manner.

Several Ways of Intercellular Communication

Chemical Messengers Gap Junctions Cell Surface Proteins Intercellular Communication by Electrical Impulses

Several Ways of Intercellular Communication

Chemical Messengers:
Cells send out signals in the form of specific chemical messengers and the target cells receive these signals and transmits into biochemical reactions. Signaling cells can simultaneously influence many cells by chemical messengers so as to enable a coordinated reaction in an organism.

Gap Junctions:
Communication between bordering cells is possible via direct contact in the form of gap junctions. Gap junctions are channels that connect two neighboring cells to allow a direct exchange of metabolites, signaling molecules and other molecules between the cells.

Cell-cell interaction via cell surface proteins:

Direct communication between cells can occur with the help of cell surface proteins. In this process a cell surface protein of one cell binds to a specific complementary protein on another cell. These bindings activate specific intracellular signaling cascade which eventually initiates specific biochemical reactions in the participating cells.

Principal Mechanisms of Intercellular Communication

Chemical Messengers Gap-Junction Cell Surface Proteins

Schematic Diagram of Intercellular Communication

Chemical Messengers
Local Chemical Messengers such as growth Factor- TGF- Beta, (Paracrine and Autocrine). Neurotransmitters (Dopamine, NO) Hormones (these are long-range chemical messengers secreted into blood by endocrine gland. Neurohormones (Vasopressin)

Several steps of intercellular communication

Receptor and Signal Transduction

Receptors (radar) are proteins. containing a binding site specific for a single chemical messenger and another binding site involved in transmitting the message. The second binding site may interact with another protein or with DNA (for steroid hormone). Type: Plasma membrane (Transmembrane) receptors Nuclear receptors

Transmembrane membrane receptor: Transmembrane receptors are a class of protein which span the plasma membrane by transmembrane domain and contain an extracellular binding domain for receiving the messenger and contain a cytosolic domain for transducing the message. Nuclear receptors: Nuclear receptors are a class of proteins found within cells that are responsible for sensing steroid and thyroid hormones and certain other molecules. Nuclear receptors have the ability to directly bind to DNA and regulate the expression of adjacent genes, hence these receptors are classified as transcription factors.

Several categories of Transmembrane membrane receptor:

Chemically gated receptors or Ion channel receptors Receptor enzyme kinases (Tyrosine kinase receptor and Serine-threonine kinase receptors). G-protein coupled receptors

Chemically gated receptors or Ion channel receptors

Receptor Enzyme Kinases

Receptor Enzyme

G-protein coupled receptors

Nuclear receptors
Receptor

Signaling via Transmembrane Receptors

Transmembrane receptors are proteins that span the phospholipid bilayer of the cell membrane. The signaling molecule (such as chemical messenger) binds on the extracellular side to the receptor. This binding activates receptor protein. Activated receptor proteins transduce the signal to the effector protein, the next component of the signal transmission pathway on the inner side of the cell membrane. In this process enzymatic activities can be triggered and/or the activated receptor engages in specific interactions with downstream signal proteins.

Structural Principles of Transmembrane Receptors

The Extracellular Domain of Transmembrane Receptors

The Transmembrane Domain The Intracellular Domain of Membrane Receptors

Regulation of Receptor Activity

Structural principles of transmembrane receptors.

Representation of the most important functional domains of transmembrane receptors

Structural principles of transmembrane receptors.

Examples of subunit structures. Transmembrane receptors can exist in a monomeric form (1), dimeric form (2) and as higher oligomers (3,4). Further subunits may associate at theextracellular and cytosolic domains, via disulfide bridges (3) or via non-covalent interactions (4).

Structural principles of transmembrane receptors.

Examples of structures of the transmembrane domains of receptors. The transmembrane domain may be composed of an a-helix (1) or several a-helices linked by loops at the cytosolic and extracellular side (2). The 7-helix transmembrane receptors are a frequently occurring receptor type. Several subunits of a transmembrane protein may associate into an oligomeric structure (3),

The Extracellular Domain of Transmembrane Receptors

The extracellular domain often contains the ligand-binding site. Glycosylation sites, i.e. attachment sites for carbohydrate residues, are also located nearby in the extracellular domain. The extracellular localized protein portion may be formed from a continuous protein chain and may include several hundred amino acids. If the receptor crosses the membrane with several transmembrane segments, the extracellular domain is formed from several loops of the protein chain that may be linked by disulfide bridges. The structure of the extracellular domain can be very diverse and is determined by the number of transmembrane sections as well as the subunit structure of the receptor.

The Transmembrane Domain The transmembrane domains have different functions, according to the type of receptor. For ligand-controlled receptors, the function of the transmembrane domain is to pass the signal on to the cytosolic domain of the receptor. For ligand-controlled ion channels, the transmembrane portion forms an ion pore that allows selective and regulated passage of ions. The transmembrane receptors span the 5nm thick phospholipid bilayer of the cell membrane with structural portions known as transmembrane elements. the transmembrane elements include 2030 mostly hydrophobic amino acids.

The Transmembrane Domain

Structure of Transmembrane Elements
High-resolution structural information about the transmembrane elements of transmembrane receptors could recently be obtained on the example of rhodopsin, the light-activated G protein coupled receptor of the vision process. These data, together with earlier data on the structures of other transmembrane proteins (e.g., bacteriorhodopsin), have confirmed that a-helices are the principal structural building blocks of the transmembrane elements of membrane receptors.

Three-dimensional structure of rhodopsin. Two views of rhodopsin. A) The seven ahelices of the G protein-coupled receptor rhodopsin weave back and forth through the membrane lipid bilayer (yellow lines) from the extracellular environment (bottom) to the cytoplasm (top). The chromophore 11-cis retinal (yellow) is nested among the transmembrane helices .B) View into the membrane plane from the cytoplasmic side of the membrane. Roman numerals indicate numbered helices.

Structure of Transmembrane Elements In addition to a-helices, proteins also use b-structures to cross the membrane. The transmembrane domain of the bacterial OmpF porin is made up of b-elements .The b-elements, in this case, are not mostly made up of hydrophobic amino acids and form a barrel-like structure.

The OmpF porin from Eschericia coli is an integral membrane channel-forming protein which spans the outer membrane in Gram-negative bacteria. The structure of a monomer of the OmpF porin is shown. In total, 16 b-bands are configured in the form of a cylinder and form the walls of a pore through which selective passage of ions takes place.

The Intracellular Domain of Membrane Receptors Two basic mechanisms are used for conduction of the signal to the inner side of the membrane. 1. * Via specific protein-protein interactions, the next protein component in the signal transmission pathway, the effector protein, is activated. * The conformational change that accompanies the perception of the signal by the receptor creates a new interaction surface for proteins that are located downstream of the receptor. * In the absence of the signal, this interaction surface is not available. Signal transmission therefore strictly depends on signal perception by the receptor, and activation of the effector molecule must be preceded by activation of the receptor by a signal.v

The Intracellular Domain of Membrane Receptors 2. * Arrival of the signal triggers enzyme activity in the cytosolic domain of the receptor. * The enzyme activity of the cytosolic domain is often tyrosine kinase activity; however, there are other examples where tyrosine phosphatase or Ser/Thrspecific protein kinase activity is activated. * In all these examples, the cytoplasmic domain carries an enzyme activity regulated by ligand binding. * The enzyme activity may be an integral part of the receptor, or it may also be a separate enzyme associated with the receptor on the inner side of the membrane.

General functions of transmembrane receptors

General functions of transmembrane receptors. Extracellular signals convert the transmembrane receptor from the inactive form R to the active form R*. The activated receptor transmits the signal to effector proteins next in the reaction sequence. Important effector reactions are the activation of heterotrimeric G-proteins, of protein tyrosine kinases and of protein tyrosine phosphatases. The tyrosine kinases and tyrosine phosphatases may be an intrinsic part of the receptor or they may be associated with the receptor. The activated receptor may also include adaptor proteins in the signaling pathway or it may induce opening of ion channels.

Regulation of Receptor Activity

These are protein sequences that permit phosphorylation of the receptor by protein kinases. Phosphorylation at Ser/Thr or Tyr residues of the cytosolic domain may lead to activation of the receptor and thus strengthen signal transmission. In this way, the protein kinases involved are often part of other signaling pathways and can link the activity of the transmembrane receptors to other signaling networks.

G protein coupled receptors

Interact with G proteins. Has 7 transmembrane receptors(7TM).
Single largest super family of proteins encoded by animal genome.

G PROTEIN COOUPLED RECEPTORS

Topology
Amino terminus faces outside of the cell Carboxy terminus faces inside of the cell Has 7 -helices

Signal transduction by GPCRs

Activation cycle of a G-protein by a G-proteincoupled receptor receiving a ligand.

G PROTEIN COOUPLED RECEPTORS