1 - Basic Concepts in Metabolism Flashcards
Typical Enzymes
-
Not a Drug Target:
-
Lysosyme
- hydrolyzes bacterial cell wall
-
Lysosyme
-
Drug Targets
-
Lactate Dehydrogenase
- Lactate -> Pyruvate
-
Carbonic Anhydrase
- CO2 -> Bicarbonate
-
Orotidine Decarboxylase
- Greatest Rate of ACCELERATION/enhancement
-
Lactate Dehydrogenase
6 Main Enzyme Classes
- Oxidoreductases
- Transferases
- Hydrolases
- Lyases
- Isomerases
- Ligases
Oxidoreductases
Redox Reactions
Lactate dehydrogenase
Transferases
Move Chemical Group
Creatine KINASE
Alanine aminotransferase
Hydrolases
Hydrolysis
Bond cleavage w/ xfer of
fxn group –> Water
Lysosyme
Chymotrypsin
Lyases
Non-hydrolytic bond cleavage
Addition of groups across a DOUBLE BOND
Fumerase
Isomerases
Intramolecular group transfer
–> change in spatial geometry
Triose Phospate isomerase
Methylmalonyl CoA mutase
Ligases
formation of New covalent bond
between substrates using NTP Hydrolysis
RNA polymerase
Pyruvate carboxylase
Enzyme Sub-Classes
Used to describe the different
Enzyme-catalyzed rxns
Amoung the PROTEINS catalyzing them
Isozyme
Catalyze the SAME rxn but differ in AA Sequence
Allows for FLEXIBILITY & CONTROl
-
Differ in catalytic activity / cofactor / regulation
- Encoded by diff genes
- Expressed at different stages of development / tissue
- Across components
-
Lactate Dehydrogenase
- 5 different Isozymes
*
- 5 different Isozymes
Cofactors
Small organic/inorganic molecule or ions
to ASSIST IN CATALYSIS
-
Dietary intake of vitamins & Minerals (IRON)
- __most can be synthesized by humans
- tetrapyrrole ring of heme groups
- but many need to be chemically modified first
- __most can be synthesized by humans
CoEnzymes
Type of Cofactor
- Held LOOSELY by enzymes
- Rxn –> Release –> Recycled (by a different set of enzymes)
- NADH / CoA
Prosthetic Groups
Type of Cofactor
- Held “TIGHTLY” by enzymes
- Recycled when IN PLACE
- Heme / Flavin / Fe-S centers
Catabolic Pathway
Break DOWN complex precursors –> simple products
Make Energy
Can occur in the same or different place as anabolism
- Proteins -> AA
- FA -> Acetyl-CoA
- Carbs -> CO2 + H2O
Anabolic / Biosynthetic
BUILD UP –> Complex Products
USE ENERGY
Can occur in the same or different place as catabolism
- AA -> Protein
- Acetyl-CoA -> Lipids
Glycolysis
Glucose -> Pyruvate
Cytosol
Fatty Acid Oxidation
FFA -> Acetyl-CoA
MITO
Amino Acid Catabolism
AA -> Ketoacids + Urea
MITO
TCA Cycle
Pyruvate -> Acetyl-CoA -> NADPH
Mitochondrial MATRIX
Oxidative Phosphorylation
ATP Synthase
NADH -> ATP
Mitochondrial MEMBRANE
Urea Cycle
AA -> Ammonia -> UREA
Cytosol->MITO
Glycogenesis
Glycogen Synthesis
Glucose –> GLYCOGEN
CYTO of muscle/liver/adipose
Glycogenolysis
Glycogen Breakdown
Glycogen -> GLUCOSE
CYTO of muscle/liver/adipose
Gluconeogenesis
AA / TG / Glycerol -> GLUCOSE
Production of GLUCOSE by non-carbohydrates
Cytosol + Mitochondria
Lipogenesis
Fatty Acid Synthesis
Acetyl-CoA –> FFA
CYTOSOL
Triglyceride Synthesis
FFA + Glycerol –> TG’s
Cytosol
Amino Acid Synthesis
Nitrogenous Molecoles -> AA
Mitochondria
Porphyrin Synthesis
Heme Synthesis
Glycine + Succinyl-CoA -> HEME
Mito + Cyto
Mevalonate Pathway
Cholesterol + Steroid Synthesis
Acetyl-CoA -> Cholesterol
CYTOSOL
Nucleotide Synthesis
Phosphate + Sugar + N-Base -> NUCLEOTIDE
Both Cyto/mito
Catabolic Funnel Stages
- Complex Molecules -> Monomers
- Fats / Polysacchrides / Proteins
-
Monomers -> Simple Intermediates
- –> Acetyl-CoA
-
Produce Energy
-
TCA Cycle + Oxidative phosporylation
- Recycles cofactors
-
TCA Cycle + Oxidative phosporylation
Committing Step
First Unique Step
Typically the natural choice for a point of REGULATION
PFK-1 = first unique step in Glycolysis
How to decide on which step we should regulate
- Target the MOST EFFICIENT SITE (typically most unique)
-
Junctions of metabolic/signaling paths
- Not just the start or the unique step
-
Glycolysis - G6K step is also used in
- spinal/sensory neurons outide of the cell
-
Junctions of metabolic/signaling paths
PFK-1
Committing Step of Glycolysis
NOT used in any other pathways
F6K -> F1,6Bisphosphate
Sites of Glycolysis Regulation
PFK-1 = commiting step
HEXOKINASE (Start)
Glucose -> G6P
PYRUVATE KINASE (end)
-> Pyruvate
3 (6) Basic Ideas for
Maintaining Metabolic Homeostasis
- Feedback
- Crossover Theorem
- Energy Charge
- Compartmentalization
- Tissue Communication / signal transduction
- Energy Regulation
Tisssue Communication / Signal Transduction
Maintaining Metabolic Homeostasis
Hormones / Cytokines / Nervous System
used to Maintain Metabolic Homeostasis
Feedback
Maintaining Metabolic Homeostasis
Returns a portion of the Output of a system as an INPUT
Typically is NEGATIVE FEEDBACK
Negative Feedback
Returns the system to the STATUS QUO
MAINTAIN HOMEOSTASIS
ATP -/-> Glycolysis
UTP + CTP -/-> Pyrimidine synthesis
Contraction of iris in eye in bright light
Positive Feedback
Can be DANGEROUS -> Runaway reactions
relatively rare in metabolism
Microphone shriek
Cancer
Crossover Theorem
- Activating or inhibiting an enzyme in a pathway will result in:
- Changes in the metabolite concentrations before & after that step
- Changes occur in opposite directions
-
PKU
- MORE PKU -> Drop in TYR production
- but also a rise in the metabolites of PHE
- MORE PKU -> Drop in TYR production
Phenylalanine Hydroxylase
Phe + BH4 -> Tyr
BH4 is a COFACTOR for the enzyme
Dihydrobiopterin Reductase
BH2 + NADPH -> BH4
BH4 is a needed cofactor for Phenylalanine Hydroxylase
( Phe -> Tyr )
Phenylketouria
PKU
Lack of Phenylalanine Hydroxylase
Phe -/-> Tyr
-
Tyrosine becomes Essential
- used for Catacholamines (hormones + NTs)
- -> Mental Retardation / Neurological Upsets
- used for Catacholamines (hormones + NTs)
-
Build up of Phe & its unusual metabolites
- “Cross over Theorem”
- phenylacetate
Varient PKU
Defects in metabolism of BH4
Treated with BH4 supplement
BH4 needed for Phenylalanine Hydroxylase
+ L-Dopa & 5 hydroxytryptophan for NT synthesis
PKU Diagnosis + Treatment
Lab Test on blood / Urine (Mass Spec)
- Dietary Interventions:
- Supplement Tyrosine
- Restrict intake of Phe
- Avoid protein rich foods, Aspartame
HMG-CoA Reductase
Comitting step in Cholesterol Synthesis
-
Statin Inhibition exploits the Cross Over Theorem
- Leads to less cholesterol
- HMG-CoA -> Mevalonic Acid ->->-> Cholesterol
Energy Charge
EC
Maintaining Metabolic Homeostasis
Ratio of Available Phosphoanhydrade Linkages
ATP + 1/2 ADP
—————-
ATP + ADP + AMP
Entire pool of adenine nucleotides
Higher when MORE ENERGY is available for Work
Energy Charge Graph
-
Steepest Parts = Most sensative to changes in EC
- 0.80 - 0.95 for most cells
- HIGH EC
- inhibits pathways GENERATING ENERGY
- STIMULATES pathways that consume energy
Glycolysis Regulation @ PFK-1
-
Stimulated by:
-
ADP / AMP / cAMP / Fructose 2,6-bisP
- PFK-2 forms F26BP but is not in glycolysis
- This is a rare example of POSITIVE FEEDBACK
-
ADP / AMP / cAMP / Fructose 2,6-bisP
-
Inhibited by:
-
ATP & Citrate
- NEGATIVE FEEDBACK
-
ATP & Citrate
Compatmentalization
Maintaining Metabolic Homeostasis
Major OPPOSING pathways are often located in DIFFERENT intracellular compartments
membrane barriers / special transport systems
- Compartments allow for control of:
- Concentrations
- Seperation of competting processes
- Fatty Acid Oxidation in MITO____
- Fatty Acid Syntheis in CYTO
Regulation of Enzymes
Maintaining Metabolic Homeostasis
Control Concentration of Materials W/
Enzymes / Substrates / Cofactors / Products
Isozymes for flexibility
Enzymes Activation/deactivation
Covalent Modification
Ways to Activate / Deactivate Enzymes
Often used in Signaling Cascades
Can be Reversible or Not
- Phosphorylation
- Acetylation
- Proteolytic Processing
- Glycosylation
- Adenylation
- Methylation
Reversible Non-Covalent Modifications
Ways to Activate / Deactivate Enzymes
Allosteric Effectors
Often the effector concentation is connected to overall metabolic state of cell
Phosphorylation
Covalent Modification
Sensitive to EC
Phosphate -> OH
of Ser Thr Tyr
Acetylation / Acylation
Covalent Modification
Sensitive to Metabolic State of Cell
Acetyl -> OH
of Ser / Thr
Fatty Acid (Acyl) -> Direct protein
like inserrtion in a membrane
Proteolysis / Proteolytic Cleavage
Covalent Modification
Activate Proenzymes / Pro-hormones
Deactivate / Recycle Mature proteins / peptides / enzymes
INSULIN needs to be proteolytically activated
Glycosylation
Covalent Modification
Sugar -> Protein Site
Used in TARGETING the protein to a cell location
or in PROTECTION in cleavage / denaturing agents
Allosterism
Change Shape & Activity
R Form = Active
-
ATCase
- key enzyme in PYRIMIDINE Synthesis
- responds to effectors & contraolled by allosterism
-
Hemogloblin
- Oxygen transporter
- Changes shape upon Oxygen uptake/release
R Form
Allosterism
Active & Open
conformation
T Form
Allosterism
LESS Active
conformation
Reciprocal Regulation
Cell will contain enzymes for both
Synthysizing & Breaking down
the same set of biochemical compounds
- Leads to PROBLEMS if in SAME COMPARTMENT
- Waste of energy / resources
- Loss of time
- –> subcellular organelles w/ specialization
How to Achieve Homeostasis
- Use Two of the main control mechanisms
- Covalent Modification
- NonCovalent binding of small efector molecules
-
To alter activity
- Can be employed at the SAME TIME
- –> Regulate a PAIR of competing pathways
- Ex.
- Glycogen synthesis & breakdown
- AA synth/breakdown