Oxidarea AG

Oxidarea glicerolului Sinteza corpilor cetonici

obiectivele
Oxidarea acizilor grai: a) saturai cu numr par de atomi de carbon; b) nesaturai cu numr par de atomi de carbon; c) saturai cu numr impar de atomi de carbon; d) n peroxizomi. Reaciile pariale, enzimele, coenzimele, reglarea, randamentul energetic. Catabolismul triacilglicerolilor reaciile pariale, enzimele, reglarea. Oxidarea glicerolului reaciile pariale, enzimele, coenzimale, reglarea, randamentul energetic al oxidrii anaerobe i aerobe. Oxidarea fosfo-, sfingo- i glicolipidelor. Metabolismul corpilor cetonici. Cile biosintezei i utilizrii lor reaciile pariale, enzimele, coenzimele, reglarea. Rolul biologic al corpilor cetonici.

Oxidarea AG saturai cu numr par de atomi de carbon

1.

2.
3.

oxidarea AG (degradarea, scindarea, catabolizarea oxidativ a AG) moleculele de AG sufer un atac oxidativ n poziia , urmat de desprinderea unui fragment cu 2C (Acetil Co A) 3 etape: Activarea AG (citoplasm) Transferul lui Acil CoA n mitocondrii b oxidarea propriu zis (mitocondrii)

Activarea AG:

R-COOH + ATP R-COO-AMP + PPi aciladenilat PPi 2 Pi R-COO-AMP + HS-CoA R-CO~SCoA + AMP
acil-CoA

Sumar:
R-COOH + ATP + HS-CoA R-CO~SCoA + AMP + PPi
R-COOH + ATP + HS-CoA +H2O R-CO~SCoA + AMP + 2 Pi E- acil Co A sintetaza Activatori: K; Mg Inhibitori: Na ; Li

Transferul lui Acil CoA n mitocondrii

Acil CoA nu poate penetra membrana intern a MC Este transportat cu ajutorul carnitinei (-hidroxi-trimetilaminobutirat), ce se formeaz din Lyz i Met activ cu participarea vitaminei C, B6, NAD

Transferul lui Acil CoA n mitocondrii

b oxidarea propriu zis

1. 2.

3.
4.

Localizat n MC repetarea a 4 reacii: Dehidrogenarea lui acil Co A (FAD) hidratare a doua dehidrogenare (NAD) reacie tiolazic n rezultat - se formeaz acetil CoA i acil CoA cu doi atomi de carbon mai puin

Bilanul energetic
Stoichiometria unui ciclu de oxidare: CH3- (CH2)n-CH2 CH2-COSCoA +FAD+ H2O+NAD+HSCoA Acil CoA (Cn-2) +FADH2+NADH+H+ Acetil CoA Stoichiometria oxidrii a. palmitic (C16): n/2 -1 numrul de cicluri pn la oxidarea complet n numrul atomilor de C

Stoichiometria oxidrii a. palmitic

16/2 -1 = 7 cicluri 7FADH2-------- 7 X 2=14ATP 7NADH+H ----- 7X3=21 ATP 8 CH3COSCoA--- 8X12= 96 ATP Sumar: 131 mol de ATP Deoarece 2 legturi macroergice sunt irosite la activarea acidului beneficiul net este de 129

Oxidarea AG nesaturai

b-oxidarea AG nesaturai se desfoar normal pn n vecintatea legturii duble (cis configuraie) Dup trei cicluri normale de b-oxidare se ajunge la un cis 3 enoil CoA. Sub aciunea izomerazei legtura dubl din cis 3 trece n trans- 2 se formeaz trans 2 enoil CoA, intermediar normal al b-oxidrii.

Exemplu: oxidarea acidului oleic (C18:19) CH3-(CH2)7-CH=CH-(CH2)7-COOH

Pentru AG polienici e necesar i o alt enzim epimeraza, care modific configuraia grupei OH la C3. Aceast E e rezultat din hidratarea legturii duble Dizomer-3 hidroxiacil CoA, ce nu poate fi substrat al enzimei de tipul L

Oxidarea AG cu numr inpar de atomi de C

Se oxideaz n acelai mod ca AG saturai, dar n ultima etap se formeaz o molecul de propionil CoA i una de Acetil CoA.

Oxidarea AG cu numr inpar de atomi de C

E- propionil CoA carboxilaza Co- vitamina H (biotin dependent) E- Metilmalonil-mutaza Co- vitamina B12 Lipsa acestei E acidemie metilmalonic (n snge i urin apare acidul metilmalonic, micornd pH sngelui (administrat vitamina B12)

Oxidarea AG n peroxisomi
Caracteristic AG C20-C26 Produsul final este Acetil CoA, dar nu este asociat cu sinteza de ATP (acetil CoA trece n mitocondrii unde este oxidat la CO2 i H2O) Difer de oxidarea mitocondrial prin reacia de oxidare a acilCoA la enoil-CoA (E- oxidaz) R-(CH2)n-COSCoA+O2 R-(CH2)-CH=CH-COSCoA + H2O2 ( sub aciunea catalazei 2H2O2 2H2O+O2) Amploarea acestui proces variaz n dependen de factorii nutriionali, hormonali, medicamentoi. Numrul peroxisomilor crete n diabet, inaniie, la administrarea unor medicamente (aspirina, preparate hipolemiante) Absena peroxisomilor- sindromul Zellweger: creterea AG cu catena lung i deces n primele luni de via

Oxidarea
1.

2.

Predomin n esutul nervos (creier) Se formeaz hidroxiacizii grai superiori, proprii lipidelor SNC Necesit: NAD, Vitamina C, ATP, O2, Fe2+ Nu intervine CoA i nu se formeaz ATP E- acid gras peroxidaza (necesit H2O2, ce rezult prin autooxidarea flavinenzimelor) Au loc concomitent 2 procese: eliminarea carboxilului sub form de CO2 oxidarea lui C la aldehid Aldehida poate fi redus la alcool sau oxidat la acidul corespunztor Nu are loc degradarea total a AG, deoarece E este activ numai la AG C13-C18.

Oxidarea

Are loc n microsomi Necesit: O2, NADPH, citocromul P450 E monooxigenaza hepatic AG se degradeaz n final prin beta oxidare

Metabolismul TG
1.

2.

n plasm exist 2 fluxuri de TG: CM transport TG exogene de la intestin la esuturi VLDL transport TG endogene- de la ficat spre esuturi Mobilizarea TG din esutul adipos are loc n etape, pn la glicerol i AG, sub aciunea lipazelor (mono-; di- , triacilglicerollipaza).

Soarta AG i glicerolului:
AG sunt transportai spre esuturi de albumina seric, unde: 1. se supun b oxidrii ( pentru a obine ATP) sau acetil-CoA (rezultat prin b-oxidare) poate fi utilizat la sinteza Col, corpilor cetonici. 2. Se activeaz i particip la sinteza TG, depozitate n esutul adipos 3. Difuzeaz n plasm i circul sub form de AG liberi (sunt captai de esuturile periferice: muchii scheletici, miocard, rinichi, ficat) Eritrocitele i creierul nu pot utiliza AG ca surs de energie Glicerolul: 1. Sinteza de TG i FL 2. Gluconeogenez 3. Oxideaz pn la CO2 i H2O

Oxidarea glicerolului
E1 glicerolkinaza E2 glicerolfosfatDH

Trigliceridlipaza

Enzima cheie a lipolizei - trigliceridlipaza adipocitar, cunoscut ca lipaza hormonsensibil. Enzima este convertibil prin fosforilare defosforilare. Forma fosforilat este activ. Catecolaminele (adrenalina, noradrenalina) snt factori majori lipolitici. Glucagonul are acela efect. Insulina, prostoglandina E snt factori antilipolitici, ei favorizeaz sinteza de TG n esutul adipos.

Sinteza corpilor cetonici (cetogeneza)

Principala cale de metabolizare a acetil CoA includerea n ciclul Krebs (n condiiile n care scindarea lipidelor i a glucidelor este echilibrat)- lipidele ard n flacra glucidelor n lipsa glucidelor; inaniie, diabet - OA se utilizeaz pentru generarea Gl. n lipsa OA, Acetil Co A recurge la formarea corpilor cetonici:

acetoacetatul, -hidrohibutiratul i acetona

Sinteza lor are loc n ficat, dar se utilizeaz de esuturile periferice Au rol energetic (muchiul cardiac, stratul cortical al rinichilor)

Utilizarea corpilor cetonici

1.

Acetoacetatul 2 mol de acetil CoA, utilizate ulterior n ciclul Krebs (23 ATP) A doua cale de activare a acetoacetatului poate fi: Acetona:
pn la propandiol (CH3-CHOHCH2OH) , scindat la fragmente acetil i formil Transformat n piruvat (prin hidroxilare dubl)

2.

Cetonemie, cetonurie

Cetonemie- mrirea c% de corpi cetonici n snge Cetonurie apariia CC n urin Diete bogate n lipide, srace n glucide; inaniie, diabet, dereglri gastrointestinale la copii sau gravide; glucozurie renal Eliminarea hidroxibutiratului i acetoacetatului din organism (fiind anioni la excreie) conduce la pierderea de cationi Na- rezult cetoacidoza Pierderea H2O dehidratarea organismului

O H3C C SCoA

acetyl-CoA
O

OOC

CH2

SCoA

malonyl-CoA

Control of fatty acid oxidation is exerted mainly at the step of fatty acid entry into mitochondria. Malonyl-CoA (which is also a precursor for fatty acid synthesis) inhibits Carnitine Palmitoyl Transferase I.

Malonyl-CoA is produced from acetyl-CoA by the enzyme Acetyl-CoA Carboxylase.

AMP-Activated Kinase, a H3C C sensor of cellular energy acetyl-CoA levels, is allosterically ATP + HCO 3 activated by AMP, which is high in concentration when ADP + Pi [ATP] is low.

SCoA

Acetyl-CoA Carboxylase is OOC CH2 C SCoA inhibited when malonyl-CoA phosphorylated by AMPActivated Kinase, leading to decreased malonyl-CoA.

Acetyl-CoA Carboxylase (inhibited by AMP-Activated Kinase)

The decrease in malonyl-CoA concentration leads to increased activity of Carnitine Palmitoyl Transferase I.
Increased fatty acid oxidation then generates acetyl-CoA, for entry into Krebs cycle with associated ATP production.

dimethylisoalloxazine
H C H3C H3C C C C H C C N CH2 HC OH OH N C C

O C NH C N O

2e +2H

H C H3C H3C C C C H C C

H N C C N CH2 HC OH OH

C NH C N H O

FAD

HC HC H2C

OH O O P OO

O P OO

Adenine Ribose

FADH2

HC HC H2C

OH O O P OO

O P OO

Adenine Ribose

The carbonyl O of the thioester substrate is hydrogen bonded to the 2'-OH of the ribitol moiety of FAD, giving the sugar alcohol a role in positioning the substrate and increasing acidity of the substrate a-proton.

Human genetic diseases have been identified that involve mutations in:

the plasma membrane fatty acid transporter CD36 Carnitine Palmitoyltransferases I & II (required for transfer of fatty acids into mitochondria) Acyl-CoA Dehydrogenases for various chain lengths of fatty acids Hydroxyacyl-CoA Dehydrogenases for medium & short chain length fatty acids Medium Chain b-Ketothiolase the trifunctional protein complex Electron Transfer Flavoprotein (ETF).

Human genetic diseases: Symptoms vary depending on the specific genetic defect but may include: hypoglycemia and failure to increase ketone body production during fasting fatty degeneration of the liver heart and/or skeletal muscle defects maternal complications of pregnancy sudden infant death (SIDS). Hereditary deficiency of Medium Chain Acyl-CoA Dehydrogenase (MCAD), the most common genetic disease relating to fatty acid catabolism, has been linked to SIDS.

The reactions presented accomplish catabolism of a fatty acid with an even number of C atoms & no double bonds. Additional enzymes deal with catabolism of fatty acids with an odd number of C atoms or with double bonds.

The final round of b-oxidation of a fatty acid with an odd number of C atoms yields acetyl-CoA & propionyl-CoA. Propionyl-CoA is converted to the Krebs cycle intermediate succinyl-CoA, by a pathway involving vitamin B12 (to be presented later).

Most double bonds of naturally occurring fatty acids have the cis configuration.

As C atoms are removed two at a time, a double bond may end up in the wrong position or wrong configuration to be the correct substrate for Enoyl-CoA Hydratase.
The reactions that allow unsaturated fatty acids to be fully catabolized by the b-oxidation pathway are summarized in the textbook.

Glucose-6-phosphatase glucose-6-P glucose Gluconeogenesis Glycolysis pyruvate fatty acids

During fasting acetyl CoA ketone bodies or carbohydrate cholesterol starvation, oxaloacetate citrate oxaloacetate is depleted in liver Krebs Cycle due to gluconeogenesis. This impedes entry of acetyl-CoA into Krebs cycle.

Acetyl-CoA in liver mitochondria is converted then to ketone bodies, acetoacetate & b-hydroxybutyrate.