midterm

Help!

substrate

products

Cofactors, Coupled Rxn, ETC

Control Mechanism

enzymes

pyruvate (in anaerobic)

Lactate and Alanine

1. NADH ----> NAD 2.Transamination reaction between amino acid form and keto form. Alanine trasnaminates to gluamate which becomes alphaketogluterate. Nees Pyridoxal phosphate. (OTC is Pryidoxidine)

oxygen. lack of O2 causes pyruvate to form lactate and alanine.

lactate dehydrogenase

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GLUT 1

erythrocytes

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GLUT 2

liver and pancreas

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GLUT 3

brain

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GLUT 4

skeletal muscle and apipose tissue

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GLUT 5

small intestine

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what stimulates PFK1?

unphosphrylated PFK2

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Is phosphotase is phosphorylated, it is

ACTIVE

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if PFK2 is phosphorylated, it is

NOT active, not producing PFK1

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what happens to phosphotase when it is active

stays there until it's needed. Interacts with another dimer and degrades into cell.

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What typically promotes phosphoylation?

glucagon

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When ratio is high (high insulin), what now happens in regards to PFK?

low glucagon, glucose into cell, Phosphorylation does not occur, PFK 2 active, PFK1 upregulated by fructo 26 bisP

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There are PFK I and PFK II. Same substrate, different product. What does 2,6 do???

This stimulates, upregulates PFK1, otherwise you would have a toxicity of fruc 6 phosphate. (then makes sorbitol…extra glucose).So it builds up PFK I. To then continue pathway.

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when glucagon receptor is stimulated what happens?

adenolyne cyclase + ATP = cAMP, stimulating protein Kinase A, phosphorylating PFK 2

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mtDNA

Ring structure. Unchanged.

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mtRNA

Does own transcription, translation, cell division.

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in regards to PDH, the pyruvate attaches where?

on outer surface of mitochondria

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where does PDH complex take place

inner surface of outer membrane of mito

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pyruvate (PDH complex)

hydroxyethyl (from ethanolthiaminepyrophosphate)+ CO2

thiamine.(thiamine pyrophosphate), This attaches to E1 so it can accept pyruvate.

controlled by ATP/ADP and NADH/NAD ratio

pyruvate decarboylase

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What is the first vitamin associated with PDH

Thiamine (inactive form), Thiamine pyrophosphate (active form)

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hydroxyethyl

2H+ + acetyl CoA

1. CoA 2. interediates: acetyolipamide, dihydrolipamide

activated by lysil residue

dihydrolipoyltransacetalase

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what stimulates E enzymes to be reactive?

cortisol (cortocortisoids)

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Coenzyme A : active form of

panothenic acid.

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how does CoA work in E2?

□ Kicks acetyl group off of acetyolipoamide and transfers it to matrix of Mito in which TCA cycle occurs.

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NADH

Riboflavin (FAD). protons attach and make FADH2 as intermediate. Coupled rxn with NAD. ReDox. Niacin. FADH2 gives protons to NAD to form NADH.

FAD activites E3

dihydrolioplydehydrogenase

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What is the first vitamin associated with E1? What is its active form?

thiamine, thiamine pyrophosphate

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where did the hydroxyethyl in E1 and E2 comes from/. (what molecule?)

ethanolthiaminepyrophosphate (hydroxyethylthiaminepyophosphate)

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which one of the vitamins would you need to properly consume for make PDH

thiamine, lipoic acid (OTC is alpha lipoic acid), panthothenic acid, riboflavin, niacin)

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how do you reverse vitamin difeciencies in mitochondria?

build up pyruvate and lactate, reduce carbs, increase vitamins, feed with lipids

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Why feed with lipids when vitamin defeicient?

because lipid metabolism does not occur in PDH.

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The PDH is regulated by

stimulation by inhibition

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what is the first regulation of carbohydrate metabolism in Mitochondria

PDH complex

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when the E are not phosphylated are they active or inactive?

active

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What does Kinase do?

only adds Phosphates

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What enzyme adds phosphates to PDH to make it Inactive?

protein kinase

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when phosphoryated, E2 is active or inactive?

inactive

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NADH/NAD is low, there is higher proportion of NAD+, this INHIBITS or ACTIVATES protein kinase.

inhibits (lacking energy, so it inhibits pK from phosphorylating and therefore inactiving enzyme. We want it active, so we want to inhibit Pk.

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If ATP is high, NADH high. What happens in PDH?

do not inhibit enzyme, phosphorylate enzyme and make inactive (via Protein Kinase)

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What stimulates PDH other than NAD and ATP ratios?

Insulin, Norepi, epi, cortisol

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What vitamins and minerals are needed for epinephrine synthesis?

Vitamin C, Copper

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Tyrosine

norepinephrine

Vitamin C. Intermediates are 3,4 dihydroxypheylalanine (DOPA) and Dopamine, copper

cortisol in adrenal medulla

1. tyrosine hydroxlase 2. DOPA decarboxylase 3. Dopamine Beta hydroxylase

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norepinephrine

epineprine

SAM-E

cortisol in adenal medulla

Phenylethanolamine N Methyl transferase

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what does a lack of copper affect?

reduced absorbtion in the gut, formation of epinephrine, Ldopa and melanin production.

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What are the defects in PDH? (conditions)

Lactic Acidosis, Neurological disorders, high pyruvate in serum, high alanine in serum.

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How would glucogenic Amino acids be affected if there was a defect in PDH?

they would end at Pyruvate. No other pathways for them to take other than PDH

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What would happen to ketogenic AA if a defect occured in PDH

nothing. They would go onto Krebs by entering into mitochondria from another point. They do not require PDH.

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TCA cycle occurs where?

matrix of Mitochondria.

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Acetyl CoA (from E2) + Oxaloacetate

Citrate

1. exergonic condensation reaction. 2. COA-SH is liberated into matrix

Inhibitors: ATP, NADH (high energy) 2. Succinyl CoA (end result of FA breakdown) 3. therefore, activated by ADP,NAD+ and low Succinyl CoA levels.

Citrate Synthase

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Citrate

Isocitrate

Intemediate: cis-aconitate 2. both a dehydration and hyration rxn.

inhibited by flourocitrate (rat Poison)

cis-aconitase

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Isocitrate

CO2 + NADH (3 ATP)+ alpha Ketogluterate

trytophan/niacin 2. this is oxidative decarboxylation

activation: ADP, Ca+. inhibited: ATP, NADH.

isocitrate dehydrogenase

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What reaction in TCA cycle first produced ATP?

Isocitrate (reaction 3)

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alpha ketogluterate

succinyl CoA + CO2 + NADH (3ATP)

Niacin, Riboflavin, thiamine, alpha lipoic acid.

Inhibited: Succinyl CoA (feedback), NADH, ATP

alphaketogluterate dehydrogenase complex. Contains tightly bound Tpp,lipoamide, FAD.

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Succinyl CoA

Succinate + ATP

Coupled Reaction: 1. GDP + Pi --> GTP 2. ADP + Pi ---> ATP catalyzed by Nucleotidediphosphate

not a control point.

Succinate thiokinase (kinase) (synthase)

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what reaction in TCA is by substrate level phosphorylation?

Reaction 5, Succinyl CoA --> Succinate.

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What s the only step where ATP is directly produced (in TCA?)

Succinyl CoA to Succinate

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Succinate

Fumerate

FADH2 (2ATP) 2. oxidation rxn

Iron in porphyrin ring facilitates rxn. Covalently binds to FAD. Inhibited by Copper and Zinc???

succinate dehydrogenase

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Fumerate

Malate

Hydration Rxn, isoergonic.

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Fumerase

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Malate

Oxaloacetate + NADH (3ATP)

NAD (niacin). Slightly endergonic (favors Malate)

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malate dehydrogenase

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How much energy produced from TCA cycle? What steps?

12 total per round!. 3 from isocitrate dH, 3 from alphaketoglut DH, 1 from succinate synthase, 2 from succinate DH, 3 from malate DH

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How many ATP produced in the entire mitochondria from one mole of glucose?

3 (PDH) + 12(TCA) times 2 (6 carbon) ---> 30 total in mitochondria.

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how many theo ATP from one mole o glucose in TCA cycle?

24 (12 x2)

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how many moles of CO2 produced in Mitochondria. Where from? (overall)

2 in PDH (E1) + 4 in TCA. Total 6 per one mole of glucose.

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What is Alphaketo form of gluterate?

alphaketogluterate

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Alpha keto form of Aspertate?

Oxaloacetate

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Alpha keto form of alanine?

Pyruvate

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Amphibolic

involved in catabolic and anabolic processes

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Anapleurotic Reactions

Increase concentrations of TCA cycle intermediates.

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IS NAD in cytosol? Does it always produce ATP?

Yes, No

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How do we pull NAD from cytosol to form ATP?

Malate Apartate Shuttle and glycertae phosphate shuttle

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Which shuttle, Malate or Glyerate would be used if energy is plentiful? Why?

Glycerate. Produces less energy than malate.

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Malate Shuttle: NADH + Oxaloacetate in Cytosol.

Malate

pyrodoxial phosphate

NADH cytosolic level

Malate dehydrogenase

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Malate shuttle: Malate inside mito

Oxaloacetate + NADH

pyrodoxial phosphate

malate dehydrogenase

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Malate Shuttle: Oxaloacetate inside Mito

Aspartate

glutamate ----> Alphaketogluterate via aminotranspherase

Aminotranspherase

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Malate Shuttle: aspartate in cytosol

Oxaloacetate

alphaketoglurate --> glutamate

Aminotransferase

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Glycerate Shuttle: dihydroxyacetone phosphate + NADH in Cytosol

Glycerol 3 phosphate + NAD+

pyrodoxial phosphate?

Glycerol 3 phosphate dehydrogenase

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Glycerate shuttle: Glycerol 3 phosphate in Mito

Dihydroxuacetone phosphate

FAD ----> FADH2

Glycerol 3 phosphate dehydrogenase

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Explain the fate of NAD in ETS (general overview)

NAD enters at complex 1. H+ go to complex 3 to form ATP, then to complex 4 to produce ATP, then to complex 5 to produce ATP.

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Explain the fate of FAD in ETS (general overview)

Enters at complex 2 and goes to 3 and 4 and 5. Produces 2 ATP

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What complex in ETS makes water? What elements are involved?

complex 4. Copper and Iron.

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Enzymes of Complex 1 in ETS

NADH dehydrogenase or NADH coenzyme Q reductase

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Complex 2 in ETS enzymes

Succinate Dehydrogenase or succinate coenzyme Q reductase

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Complex 3 enzymes in ETS

Cytochrome C - coenzyme Q oxidoreductase

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Complex 4 enzymes for ETS

Cytochrome oxidase (copper)

Copper

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Complex 5 enzymes for ETS

ATP synthase

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Inhibitors of Complex 1 ETS

insecticides, barbituates, antibiotics.

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NAD in ETS

Coenzyme Q10 (ubiquinone)

FMN (riboflavin)

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describe struture of complex 5

see notes for diagram. alpha and beta sections, concentration gradient, H+ pass through F0 and F1 to stimulate Pi + ADP ---> ATP. Turns as H+ pass.

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coupling

Whenever a H+ is going through the Core (the Gamma) of the ATP synthetase.

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un coupling

When H+ is diverted away from Gamma Region. H+ are produced, but won't produce ATP.

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inhibitors of complex 2

antibiotics and insecticides.

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inhibitors of complex 3

antibiotics (some)

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Inhibitors of complex 4

H2S, Cyanide, Azide, CO

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what vitamins are associated with ETS?

Riboflavin (FMN and FAD), Niacin

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high insulin in blood: What happens to PFK2?

1. glu into cell. 2. Fruc 6 Phosphate ---> (ATP + Mg) Fruc 2,6 bisphosphate. 3. Stimulates PFK1 in glycolitic pathway so breakdown of glu can continue. When modulated, PFK2 slows production of fructo 26 bP and pathways slows.

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when glucagon binds to cells of mitochondrial membane, what is stimulated?

Adenolyne cyclase and ATP to form CAMP. This turns on Protein Kinase A, whch stimulates with ATP to phosphorylate PFK2. PFK2 is now inhibited and breakdown of glucose is slowed.

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active form of panothenic acid?

Coenzyme A

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how many ATP and ow many CO2 are produced in PDH complex?

1 NADH (3 ATP) and 1 CO2. For glucose mole, would be DOUBLE!

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If ATP is high, then Ca+ is ______?

LOW

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What does Ca+ stimulate in PDH?

phosphotases to come off the PDH complex to make ACTIVE to get ATP production. ATP low, Ca+ high….remove PH makes it active.

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