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    Carbohydrate Metabolism

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    Yulliza Kurniawaty L
    The document discusses carbohydrate metabolism, specifically glycogen metabolism. It covers: 1) The breakdown and synthesis of glycogen, which involves glycogen phosphorylase, glycogen debranching enzyme, phosphoglucomutase, glycogen synthase, and glycogen branching enzyme. 2) Control mechanisms that ensure glycogen is either synthesized or broken down according to cellular needs, including allosteric regulation and covalent modification through cyclic cascades. 3) The role of glycogen in storing glucose in the liver and muscles, and maintaining blood glucose levels.

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    Carbohydrate Metabolism

    Uploaded by

    Yulliza Kurniawaty L
    149 views57 pages
    The document discusses carbohydrate metabolism, specifically glycogen metabolism. It covers: 1) The breakdown and synthesis of glycogen, which involves glycogen phosphorylase, glycogen debranching enzyme, phosphoglucomutase, glycogen synthase, and glycogen branching enzyme. 2) Control mechanisms that ensure glycogen is either synthesized or broken down according to cellular needs, including allosteric regulation and covalent modification through cyclic cascades. 3) The role of glycogen in storing glucose in the liver and muscles, and maintaining blood glucose levels.

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    The document discusses carbohydrate metabolism, specifically glycogen metabolism. It covers: 1) The breakdown and synthesis of glycogen, which involves glycogen phosphorylase, glycogen debranching enzyme, phosphoglucomutase, glycogen synthase, and glycogen branching enzyme. 2) Control mechanisms that ensure glycogen is either synthesized or broken down according to cellular needs, including allosteric regulation and covalent modification through cyclic cascades. 3) The role of glycogen in storing glucose in the liver and muscles, and maintaining blood glucose levels.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PPT, PDF, TXT or read online from Scribd
    Download as ppt, pdf, or txt
    149 views57 pages

    Carbohydrate Metabolism

    Uploaded by

    Yulliza Kurniawaty L
    The document discusses carbohydrate metabolism, specifically glycogen metabolism. It covers: 1) The breakdown and synthesis of glycogen, which involves glycogen phosphorylase, glycogen debranching enzyme, phosphoglucomutase, glycogen synthase, and glycogen branching enzyme. 2) Control mechanisms that ensure glycogen is either synthesized or broken down according to cellular needs, including allosteric regulation and covalent modification through cyclic cascades. 3) The role of glycogen in storing glucose in the liver and muscles, and maintaining blood glucose levels.

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    Attribution Non-Commercial (BY-NC)
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    Download as ppt, pdf, or txt
    of 57

    Carbohydrate Metabolism

    Budi Satrio
    Biochemistry Department - Medicine Faculty Al-Zaytun University Indonesia

    Standard Questions 1
    1.

    2.

    3.

    4.

    What are the major types of metabolic pathways in living organisms? What biochemical reactions are responsible for the breakdown of food molecules in energy production? How are metabolic processes regulated so that the energy and biosynthetic requirements of organisms are consistently met? How is is glycogen alternately synthesized and degraded to provide animals a consistent supply of glucose?

    Standard Questions 2
    1. 2. 3. How do cells extract energy from glucose? What is the difference between aerobic respiration and fermentation? How do glycolysis and gluconeogenesis differ? How are they similar? What regulatory mechanism ensures that glycolysis and gluconeogenesis do not occur simultaneously to any great extent?

    4.

    Major Pathways in Carbohydrate Metabolism

    The Three Stages of Catabolism

    The Glycolytic Pathway

    Glycolysis and Gluconeogenesis

    Gluconeogenesis Substrates

    The Cori Cycle

    Glucose-Alanine Cycle

    Calcium Metabolism and Blood Glucose

    Carbohydrate Metabolism

    Glycogen Metabolism 1
    I. Glycogen Breakdown II. Glycogen Synthesis III. Control of Glycogen Metabolism IV. Glycogen Storage Diseases

    Glycogen
    Glycogen serves as storage carbohydrate in animals, insects and fungi Glucose residues in a-1,4 linkage Heavily branched (a-1,6) to allow rapid mobilization of Glc from many non-reducing ends In animals, accumulated mainly in liver and muscle

    Glycogen Structure

    Glycogen plays different roles in liver and muscle


    Liver supplies tissues with Glc from glycogen during fasting In muscle, conversion of glycogen to Glc is important during strenuous exercise, and conversion of Glc to glycogen plays important role in lowering blood Glc after feeding

    Why use Glc polymer versus fatty acids for energy storage?
    Muscles cannot mobilize fat as rapidly as glycogen. Fatty acid residues of fat cannot be metabolized anaerobically. Animals cannot convert all fatty acids to Glc, so fat metabolism alone cannot adequately maintain essential blood Glc levels.

    1. Glycogen Breakdown to Glc-6-P Requires Three Enzymes


    A. Glycogen phosphorylase to cleave a1,4 linkages
    Glycogen + Pi ----------> Glycogen + Glc-1-P
    n residues n-1 residues

    B. Glycogen debranching enzyme


    a(1->4) glycosyl transferase and a(1->6) glucosidase activities -> Glc-1-P and Glc

    C. Phosphoglucomutase to convert to usable form


    Glc-1-P ----------> Glc-6-P

    Glc-6-phosphatase is required for export from liver as Glc

    A. Glycogen Phosphorylase
    Dimer of identical 97 kDa subunits

    Structural Domains and Binding Sites Each phosphorylase subunit has glycogen binding subdomain and allosteric effector binding sites and modification sites (regulatory subdomain) in the Nterminal domain. Cofactor (PLP) is linked in the Cterminal domain. The catalytic site is located at the interface of glycogen binding and regulatory subdomains with the C-terminal domain.
    Pyridoxal Phosphate is an Essential Cofactor for Phosphorylase Pyridoxal-5-phosphate (PLP), a vitamin B6 derivative, is covalently linked to phosphorylase via a Schiff base to Lys 679. In an unusual role, only the phosphate group participates in the catalytic process.

    Glycogen phosphorylase

    Glycogen phosphorylase
    Glycogen binding site is crevice on N-terminal domain Same radius of curvature as glycogen Can accommodate 4-5 Glc residues BUT no room for branches => No cleavage closer than 5 residues away from branch point

    Pyridoxal Phosphate (PLP)


    and PLP covalently bound to phosphorylase via Schiff base linkage to Phosphorylase Lys 679.

    Glycogen phosphorylase reaction mechanism


    1. Formation of an EPiglycogen ternary complex. 2. Oxonium ion intermediate (I) formation from the a-linked terminal glucosyl residue involving acid catalysis by Pi facilitated by proton transfer from phosphate of PLP. Oxonium ion I proposed because inhibition by 1,5-gluconolactone (same half-chair conformation as oxonium ion I) 3. Reaction of Pi with overall retention of configuration around C1 to form a-D-Glc-1-phosphate. The glycogen, minus one residue, cycles back to step 1.

    B. Glycogen Debranching Enzyme


    Two active sites for two different catalytic activities Required for degradation of almost half of glycogen molecule

    Maximal rate of debranching enzyme much slower than that of glycogen phosphorylase

    B. Glycogen Debranching Enzyme


    Acts as an a(14) transglycosylase (glycosyl transferase) by transferring an a(14) linked trisaccharide unit from a limit branch of glycogen to the nonreducing end of another branch. Also catalyzes the hydrolysis of the a(16) bond of the remaining glycosyl residue to yield Glc.

    C. Phosphoglucomutase
    Mechanism of action is similar to that of phosphoglycerate mutase except Ser carries phosphoryl group here.

    If Glc-1,6-bisP dissociates, enzyme is inactivated; small amounts are generated by phosphoglucokinase from Glc-1-P to prevent inactivation

    Glucose-6-Phosphatase is a Complex of Proteins

    Activity is prominent in liver, kidney as well as pancreatic islet cells, but is NOT found in muscle

    2. Glycogen Synthesis
    A. UDP-Glc synthesis

    B. Glycogen synthase
    C. Glycogenin

    D. Glycogen branching enzyme

    2A UDP-Glc Synthesis
    UDP-Glucose Pyrophosphorylase converts UTP and Glc-1-P to UDP-Glc and pyrophosphate. Inorganic pyrophosphatase converts PPi to 2Pi, driving reaction forward

    Use of UDP-Glc makes synthesis reaction favorable

    UDPGs high-energy status permits it to donate glucosyl units to the growing glycogen chain in a thermodynamically favorable reaction. This is a common strategy for carbohydrate addition.

    The overall reaction for the formation of UDPG is highly exergonic.

    UDP-Glc Pyrophosphorylase converts UTP and G1P to UDP-Glc (UDPG) and PPi. Inorganic pyrophosphatase converts PPi to 2Pi Nucleoside diphosphate kinase to replenish UTP

    UTP for the reaction is replenished through a phosphate transfer. The reaction is catalyzed by nucleoside diphosphate kinase.

    2B. Glycogen synthase


    Glc is added in a-1,4 linkage from UDP-Glc to the non-reducing end of a glycogen chain The reaction mechanism is apparently like that of phosphorylase, as the synthesis reaction also is inhibited by 1,5-gluconolactone Glycogen synthase can only extend a chaincannot initiate de novo synthesis

    2B. Glycogen Synthase

    Glycogen Synthesis

    2C. Glycogenin- a protein that primes glycogen synthesis


    The first step of glycogen synthesis is attachment of a Glc residue to Tyr194 -OH group of glycogenin by a tyrosine glucosyltransferase. Glycogenin autocatalytically extends the glucan chain by up to 7 additional UDP-Glc-supplied residues, forming a primer for the initiation of glycogen synthesis. Once primer is generated, glycogen synthase can associate to form ternary complex; upon extension of the chain, glycogenin dissociates from the complex

    2D. Glycogen branching by amylo-(1,41,6)transglycosylase. Terminal chain


    segments of about 7 residues are transferred to the C6-OH groups of Glc residues on the same or another glycogen chain. Each segment must come from a chain of >10 residues; new branch point must be at least 4 residues away from other branch points. This reflects the structure of the catalytic site.
    Donor from a branch at least 11 residues long

    New branch at least 4 residues from another branch

    Glycogen Degradation

    Glycogen Degradation

    3. Control of Glycogen Metabolism


    Due to differences in reaction, both glycogen synthesis and breakdown are exergonic under similar physiological conditions. If both pathways operate simultaneously, net result is hydrolysis of UTP. This is similar to the PFK-FBPase substrate cycle seen in glycolysis and gluconeogenesis. Glycogen phosphorylase and glycogen synthase therefore are under stringent control so that glycogen is either synthesized or utilized according to cellular needs.

    Major Factors Affecting Glycogen Metabolism

    3. Control of Glycogen Metabolism


    A. Direct Allosteric Control of Glycogen Phosphorylase and Glycogen Synthase B. Covalent Modification of Enzymes by Cyclic Cascades: Effector Signal Amplification C. Glycogen Phosphorylase Bicyclic Cascade D. Glycogen Synthase Bicyclic Cascade E. Integration of Glycogen Metabolism Control Mechanisms F. Maintenance of Blood Glucose Levels G. Response to Stress

    3A. Direct Allosteric Control of Glycogen Phosphorylase and Glycogen Synthase


    The rates of both glycogen phosphorylase and glycogen synthase reactions are under allosteric control by effectors that include ATP, Glucose-6Phosphate (G6P), and AMP. In muscle, glycogen phosphorylase is activated by AMP and inhibited by ATP and G6P. Glycogen synthase, on the other hand, is activated by G6P. When there is high demand for ATP, glycogen phosphorylase is stimulated and glycogen synthase is inhibited.

    Control of Glycogen phosphorylase activity


    Tight

    AMP

    Relaxed Less active More active

    3B. Covalent Modification of Enzymes by Cyclic Cascades: Signal Amplification


    Compared to other regulatory systems: 1. Can respond to a greater number of allosteric stimuli. 2. Exhibit greater flexibility in their control patterns. 3. Possess enormous amplification potential in their responses to variations in effector concentrations. Why? Because the enzymes that modify and demodify a target enzyme are themselves under allosteric control. It is possible for a small change in concentration of an allosteric effector of a modifying enzyme to cause a large change in the concentration of an active, modified target enzyme.

    Example 1: A monocyclic enzyme cascade


    (a)

    F = modifying enzyme
    R = demodifying enzyme e1 and e2 are allosteric effectors E = target enzyme (b)

    Small change in [e1] results in larger change in fraction of E in active form than if e1 directly regulated E

    Example 2: A bicyclic enzyme cascade


    Here, one of the modifying enzymes (F2) is also subject to chemical modification.

    Now the difference between a change in [e1] and the resulting change in fraction of active E is exponentially larger.

    3C. Glycogen Phosphorylase Bicyclic Cascade


    Allosteric control of phosphorylase allows direct control by metabolic indicators (AMP vs ATP; Glc-6-P) and bicyclic enzyme cascade adds response to hormonal signals

    Phosphorylase kinase (F2) phosphorylates glycogen phosphorylase b (E) at Ser14 -> phosphorylase a
    cAMP-dependent protein kinase (F1) phosphorylates and activates phosphorylase kinase Phosphoprotein phosphatase type 1 (PP-1 = R1 and R2) dephosphorylates both phosphorylase and phosphorylase kinase, decreasing activity

    Control of glycogen metabolism in muscle

    Cyclic AMP synthesis and metabolism


    In both the glycogen phosphorylase and glycogen synthase cascades, the primary intracellular signal is adenosine-3,5-cyclic monophosphate (cAMP).

    cAMP-Dependent Protein Kinase


    cAMP binds to the regulatory subunits (R) of cAMP dependent protein kinase (cAPK) to cause dissociation of active catalytic monomers (C).
    R2C2 + 4cAMP R2(cAMP)4 + 2C*
    cAMP2 cAMP2

    * *

    The intracellular [cAMP] determines the fraction of active cAPK and thus the rate of substrate phosphorylation.

    Catalytic (C) subunit of cAMP-dependent protein kinase in complex with ATP and 20-residue segment of a naturally occurring pseudosubstrate inhibitor (yellow).

    Phosphorylase Kinase: Coordination of Enzyme Activation with [Ca2+]


    Phosphorylase kinase has 4 nonidentical subunits forming the active oligomer, (a)4. Isolated subunits have full catalytic activity. The subunit is calmodulin (CaM), a ubiquitous Ca2+-binding regulatory protein. CaM confers Ca2+ sensitivity on the complex. When Ca2+ binds, at concentrations as low as 10-7 M, CaM undergoes a conformational change that activates phosphorylase kinase.

    Calmodulin (CaM): A Ca2+Activated Switch


    CaM has 2 high-affinity Ca2+ binding sites on each of its globular domains.

    Calmodulin
    Binding of Ca2+ to either domain of CaM induces a conformational change that exposes a buried Metrich hydrophobic patch. This patch binds to the CaM-binding domain of phosphorylase kinase subunit and modulates its activity. CaM's central alpha helix serves as a flexible tether rather than a rigid spacer.

    (Ca2+)4-CaM in complex with a 26residue target polypeptide

    Phosphorylase Kinase: Coordination of Enzyme Activation with [Ca2+]


    Muscle contraction is triggered by a transient increase in cytosolic Ca2+ released from intracellular reservoirs in response to nerve impulses. Therefore, the rate of glycogen breakdown is linked to the rate of muscle contraction to provide Glc for glycolysis and ATP production. Sites on both a and subunits of phosphorylase kinase may be phosphorylated. Phosphorylation causes the enzyme to become activated at lower concentrations of Ca2+ ; full activity is obtained in the presence of Ca2+ AND when both these subunits are phosphorylated.

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