Está en la página 1de 70

METABOLISME KARBOHIDRAT

21 NOVEMBER 2011

TUGAS
1. TULISKAN JUMLAH TOTAL ATP YANG DIHASILKAN PADA PROSES GLIKOLISIS DAN PERSAMAAN TOTAL REAKSI GLIKOLISIS (DG MENGABAIKAN ION H+)

Glycolysis - total pathway, omitting H+: glucose + 2 NAD+ + 2 ADP + 2 Pi 2 pyruvate + 2 NADH + 2 ATP Jumlah total ATP yang dihasilkan : 2 Kalau ATP dari NADH dihitung (2x3) + 2 = 8 ATP

2. Jalur glikolisis diregulasi oleh 3 enzim yang mengkatalisis reaksi-reaksi yang yg berjalan spontan, yaitu : heksokinase, fosfofruktokinase, dan piruvat kinase. Jelaskan masing-masing regulasinya !

6 CH 2OH

H
4

O H
2

ATP ADP H H
1 4

6 CH OPO 2 2 3 5

O H
2

H
1

H OH
3

OH

OH

Mg

2+

H OH
3

OH

OH

OH

Hexokinase

OH

glucose

glucose-6-phosphate

Hexokinase is inhibited by product glucose-6phosphate: by competition at the active site by allosteric interaction at a separate enzyme site. Cells trap glucose by phosphorylating it, preventing exit on glucose carriers. Product inhibition of Hexokinase ensures that cells will not continue to accumulate glucose from the blood, if [glucose-6-phosphate] within the cell is sufficient

6 CH 2OH

Glucokinase is a variant of Hexokinase found in liver.

H
4

O H
2

ATP ADP H H
1 4

6 CH OPO 2 2 3 5

O H
2

H
1

H OH
3

OH

OH

Mg2+ OH

H OH
3

OH

OH

Hexokinase

OH

glucose

glucose-6-phosphate

Glucokinase has a high KM for glucose. It is active only at high [glucose].


Glucokinase is not subject to product inhibition by glucose-6-phosphate. Liver will take up & phosphorylate glucose even when liver [glucose-6phosphate] is high.

Phosphofructokinase
6 CH OPO 2 2 3

1CH2OH

6 CH OPO 2 2 3

ATP ADP HO H
2 5

1CH2OPO32

H
4

H
4

HO H

3 OH

Mg2+

3 OH

OH

OH

fructose-6-phosphate

fructose-1,6-bisphosphate

Phosphofructokinase is usually the ratelimiting step of the Glycolysis pathway.

Phosphofructokinase is allosterically inhibited by ATP.


At low concentration, the substrate ATP binds only at the active site. At high concentration, ATP binds also at a low-affinity regulatory site, promoting the tense conformation.

Inhibition of Phosphofructokinase when [ATP] is high prevents breakdown of glucose in a pathway whose main role is to make ATP. It is more useful to the cell to store glucose as glycogen when ATP is plentiful.

Pyruvate Kinase
O C 1 C 2 OPO32
3 CH 2

ADP ATP

O
1 2

O C C O

3 CH 3

phosphoenolpyruvate

pyruvate

Pyruvate Kinase, the last step Glycolysis, is controlled in liver partly by modulation of the amount of enzyme.

High [glucose] within liver cells causes a transcription factor carbohydrate responsive element binding protein (ChREBP) to be transferred into the nucleus, where it activates transcription of the gene for Pyruvate Kinase. This facilitates converting excess glucose to pyruvate, which is metabolized to acetyl-CoA, the main precursor for synthesis of fatty acids, for long term energy storage.

KREBS CYCLE TRIVIA QUIZ

Complete the following statements: A. When 1 acetyl CoA enters the citric acid cycle, the C atoms produce ____CO2. B. In 1 cycle, a total of ____NADH are produced. C. In 1 cycle, a total of ____FADH2 are produced.
13

KREBS CYCLE TRIVIA QUIZ

Complete the following statements: A. When 1 acetyl CoA enters the citric acid cycle, the C atoms produce 2 CO2. B. In 1 cycle, a total of 3 NADH are produced. C. In 1 cycle, a total of 1 FADH2 are produced.
14

3. Jelaskan fungsi-fungsi utama (peranan) dari siklus krebs! (minimal 2)

The function of TCA cycle :


1. Oxidized of Acetyl CoA ATP ( 1 mol Acetyl CoA 12 mol ATP ) 2. The citric acid cycle is AMPHIBOLIC : - it can oxidized to yield ATP 3. It also has a central role in gluconeogenesis, lipogenesis, and interconversion of amino acids.

4. The final common pathway for the aerobic oxidation of carbohydrate, lipid and protein

The citric acid cycle is the final common pathway for the oxidation of carbohydrate, lipid, and protein because glucose, fatty acids, and most amino acids are metabolized to acetyl-CoA

ANGGOTA TCA CYCLE BERSIFAT AMFIBOLIK Dapat dioksidasi lebih lanjut menjadi energi * katabolisme asam amino anggota tca cycle energi * oksidasi beta asam lemak asetil KoA anggota siklus krebs energi * oksidasi glukosa piruvat asetil KoA anggota siklus krebs energi Dapat disintesis menjadi senyawa lain, misalnya menjadi : * glukosa (melalui glukoneogenesis) * asam amino tertentu * asam lemak (lipogenesis)

Tugas no 4 Jelaskan inhibitor-inhibitor pada siklus krebs ? (minimal 2)

INHIBITOR SIKLUS ASAM SITRAT


Fluoroasetat : * Dgn KoA-SH membentuk fluoroasetil-KoA * Fluoroasetil-KoA berkondensasi dgn oksaloasetat membentuk fluorositrat ( dikatalisis oleh sitrat sintase) * Fluorositrat menghambat enzim akonitase terjadi akumulasi sitrat * Fluoroasetat didapatkan misalnya dari pestisida Malonat : menghambat enzim suksinat dehidrogenase Arsenit : menghambat enzim -ketoglutarat dehidrogenase kompleks

VITAMINS PLAY KEY ROLES IN KREBS CYCLE


Four of the B vitamins are essential in the citric acid cycle (1) riboflavin, (VIT B2) in the form of flavin adenine dinucleotide (FAD), a cofactor for succinate dehydrogenase (2)niacin, in the form of nicotinamide adenine dinucleotide (NAD), the electron acceptor for isocitrate dehydrogenase,-ketoglutarate dehydrogenase, and malate dehydrogenase

3. thiamin (vitamin B1), as thiamin diphosphate, the coenzyme for decarboxylation in the ketoglutarate dehydrogenase reaction 4 pantothenic acid, as part of coenzyme A, the cofactor attached to "active" carboxylic acid residues such as acetyl-CoA and succinyl-CoA.

VITAMIN B1 (Thiamine) B2 (Riboflavin) B3(Pantothenate) B6(Pyridoxine) B12 (Cobalamine) Niacin=Nicotinat Folic Acid

KOENSIM TPP = Thiamine Pyrophosphate FAD = Flavin Adenine Dinucleotide CoA = Coenzyme-A PLP = Pyridoxal Phosphate 5- deoxy adenocyl cobalamine NAD = Niacin Adenine Dinucleotide Tetrahydrofolate

Biotin

Biotin

Citric Acid Cycle

Regulation of Citric Acid Cycle


Operates when ATP is needed High levels of ATP and/or NADH inhibit citrate synthetase (first step in cycle) High levels of ADP and NAD+ activate isocitrate dehydrogenase Low levels of ATP or high levels of acetyl CoA speed up the cycle to give energy

26

Summary Krebs cycle reactions

FAD

FADH2

Cellular respiration

The final stage of aerobic respiration occurs in the electron systems embedded in the inner membrane of the mitochondrion. Oxidation phosphorylation (which takes place on the cristae of the mitochondria) processes the H+ ions and electrons to generate high yields of ATP. NADH and FADH2 give up their electrons to transport (enzyme) systems embedded in the mitochondrial inner membrane.

32

Diagram of the Process


Occurs across Cristae

Occurs in Cytoplasm
Occurs in Matrix

Review of Mitochondria Structure


Smooth outer Membrane Folded inner membrane called Cristae Space inside cristae called the Matrix

definitions
Electron Transport: Electrons carried by reduced coenzymes (NADH or FADH2) are passed sequentially through a chain of proteins and coenzymes (so called electron transport chain) to O2 . Oxidative Phosphorylation: Coupling e- Transport (Oxidation) and ATP synthesis (Phosphorylation) .

Organization of Chain
NADH dehydrogenase or NADH-Q oxidoreductase (complex I) Succinate-Q reductase (complex II) Coenzyme Q (CoQ) (also called ubiquinone) Cytochrome c oxidase (complex III) Cytochrome c (Cyt c) Cytochrome c oxidase (complex IV)

Succinate-Q reductase (Complex II), in contrast with the other complexes, does not pump protons.

Each complex accepts and donates electrons to a relatively mobile electron carriers. The electrons ultimately combine with oxygen and protons to form water. It requires oxygen that is why it is also called respiratory chain. It accounts for the greatest portion of the body's use of oxygen.

Electron Transport Chain

1. 2. 3. 4. 5. 6.

Coenzymes which take part are: NAD NADP FAD FMN Coenzyme Q (CoQ) Fe-S proteins

Coenzyme Q It is also called ubiquinone. They contain isoprene units in their side chains. Can accept H atoms from both FMNH2 and FADH2. the only electron carrier not bound to a protein. it can accept/donate 1 or 2 e-. Q can mediate e- transfer between 2 e- that transfer and 1 e- carriers

When bound to special sites in respiratory complexes, CoQ can accept 1 e- to form a semiquinone radical (Q-).
O CH3O CH3 CH3 CH3O O (CH2 CH C CH3O O CH3 CH3 CH3O O (CH2 CH C CH2)nH

CH2)nH

coenzyme Q

coenzyme Q

e + 2 H+
OH CH3O CH3 CH3 CH3O OH (CH2 CH C CH2)nH

coenzyme QH2

Iron-sulfur Centers (clusters)


Iron-sulfur centers (Fe-S) are prosthetic groups containing 1-4 iron atoms
Iron-sulfur centers transfer only one electron, even if they contain two or more iron atoms. E.g., a 4-Fe center might cycle between redox states: Fe+++3, Fe++1 (oxidized) + 1 e Fe+++2, Fe++2
(reduced)

Cys S S Cys Cys S S Fe

S S Fe Fe S Fe

Fe S

S Cys Cys S S Cys

Fe S S Cys

Cys

Iron-Sulfur Centers

Complex I. Steps
1. The initial step is the binding of the NADH and transfer of 2 electrons to the FMN (Flavin mononucleotide) to give the reduced form of FMNH2 2. Electrons are then transferred from FMNH2 to a series of iron-sulfur clusters (Fe-S cluster), the second type of prosthetic group in complex I. 3. Electrons in the iron-sulfur clusters of NADH-Q oxidoreductase are shuttled to coenzyme Q.

The initial electron transfers are: NADH + H+ + FMN NAD+ + FMNH2 FMNH2 + (Fe-S)ox FMNH + (Fe-S)red + H+
After Fe-S is reoxidized by transfer of the electron to the next iron-sulfur center in the pathway: FMNH + (Fe-S)ox FMN + (Fe-S)red + H+ Electrons pass through a series of iron-sulfur centers, and are eventually transferred to coenzyme Q. Coenzyme Q accepts 2 e and picks up 2 H+ to yield the fully reduced QH2.

The flow of two electrons from NADH to coenzyme Q through NADH-Q oxidoreductase leads to the pumping of four hydrogen ions out of the matrix of the mitochondrion.

Complex II
1. Complex II accepts electrons from succinate formed during the TCA cycle. 2. Electrons flow from succinate to FAD (flavin-adenine dinucleotide) coenzyme, through an iron-sulfur protein and a cytochrome b550 protein (the number refers to the wavelength where the protein absorbs), and to coenzyme Q.

No protons are translocated by Complex II. Coenzyme Q is not bound to a protein; instead it is a mobile electron carrier and can float within the inner membrane, where it can transfer electrons from Complex I and Complex II to Complex III. Coenzyme Q can accept electrons from both FMNH (produced by complex I), and from FADH (complex II) which is produced by succinate dehydrogenase and acyl CoA dehydrogenase.

Complex III

Complex III accepts electrons from reduced coenzyme Q, moves them within the complex through two cytochromes b, an iron-sulfur protein, and cytochrome c1. Electron flow through Complex II transfers proton(s) through the membrane into the intermembrane space.

Cytochrome c transfers its electrons to the final electron transport component, Complex IV, or cytochrome oxidase.

Cytochromes
Cytochromes are electron carriers containing hemes .

Hemes in the 3 classes of cytochrome (a, b, c) differ in substituents on the porphyrin ring.
Some cytochromes(b,c1,a,a3) are part of large integral membrane protein complexes. Cytochrome c is a small, water-soluble protein.

CH3 CH3 HC S CH2 protein

N H3C N

CH3 Fe N CH3 N CH S CH2 protein

OOC

CH2 CH2

CH2 CH2 COO

CH3

Heme c

Heme is a prosthetic group of cytochromes. Heme contains an iron atom in a porphyrin ring system.
The heme iron can undergo 1 e- transition between ferric and ferrous states: Fe3+ + e- Fe2+ Copper ions besides two heme A groups (a and a3) act as electron carriers in Cyta,a3 Cu2++e- Cu+

Complex IV (cytochrome c oxidase)

Complex IV transfers electrons through a copper-containing protein, cytochrome a, and cytochrome a3, and finally to molecular oxygen.

Cytochrome a+a also called Cytochrome oxidase is the only electron carrier that has got a free ligand i.e. bound copper that can react directly with molecular oxygen and produce water.

Composition of Respiratory Chain Complexes


Complex Name No. of Proteins Prosthetic Groups

Complex I

NADH Dehydrogenase
Succinate-CoQ Reductase CoQ-cyt c Reductase Cytochrome Oxidase

46

FMN, 9 Fe-S cntrs.


FAD, cyt b560, 3 Fe-S cntrs. cyt bH, cyt bL, cyt c1, Fe-SRieske cyt a, cyt a3, CuA, CuB

Complex II

Complex III

11

Complex IV

13

The transfer of electrons is not directly to oxygen but through coenzymes


NAD+ FMN FeS FAD FeS ubiquinone Cyt b ubiquinone FeS Cyt c1 Cyt c Cyt a Cyt a3 1/2 O2

There are 2 sites of entry for electrons into the electron transport chain:

NAD+ or FAD Both are coenzymes for dehydrogenase enzymes

Mitochondrial Complexes
NAD+ FMN

I
FeS FAD FeS ubiquinone Cyt b

NADH Dehydrogenase

II
Succinate dehydrogenase
ubiquinone

Cytochrome Oxidase

FeS

Cyt c1

Cyt c

Cyt a

Cyt a3 1/2 O2

III
CoQ-cyt c Reductase

IV

H+ Transport
Complex I, III, IV drive H+ transport from matrix to the cytosol When e- flow through, which creates proton gradient(electrochemical potential) across the inner membrane Complex I and Complex IV : The mechanism of H+ transport is still not known. The mechanism of H+ transport in Complex III is Q cycle.

Matrix
H+ + NADH NAD+ + 2H+ 2H+ + O2 H2O

2 e Q

III IV

++
4H+ 4H+

cyt c

2H+

Intermembrane Space

4H+ are pumped per 2e passing through complex III. The H+/e ratio is less certain for the other complexes: probably 4H+/2e for complex I; 2H+/2e for complex IV.

Q Cycle :The mechanism of H+ transport in Complex

III

matrix Q

2 H+ Q. cyt bH
Complex III

QH2

QH2

cyt bL Q e Q.

QH2 e Fe-S 2 H+ cyt c1 cyt c

intermembrane space

1.Electrons are transported along the inner mitochondrial membrane, through a series of electron carriers 2.Protons (indicated by + charge) are translocated across the membrane, from the matrix to the intermembrane space 3.Oxygen is the terminal electron acceptor, combining with electrons and H+ ions to produce water 4. As NADH delivers more H+ and electrons into the ETS, the proton gradient increases, with H+ building up outside the inner mitochondrial membrane, and OH- inside the membrane.

The overall pathway for electron transport is therefore:

How is the oxidation of NADH coupled to the phosphorylation of ADP?