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INTRODUCTION
I. BIOMEDICAL IMPORTANCE
A. Porphyrias
B. Jaundice
II, IMPORTANCE OF METALLOPORPHYRINS AND HEMOPROTEINS
A. Porphyrins
B. Substituent Side Chains in Porphyrins
1. Type III Porphyrin
2. Type I Porphyrin
BIOSYNTHESIS OF HEME
CATABOLISM OF HEME
BIOENERGETICS
I. CONCEPT OF FREE ENERGY
II. KINDS OF CELLULAR REACTIONS
A. Exergonic Reactions
B. Endergonic Reaction
III. REDOX REACTIONS
A. Oxidation
B. Reduction
IV. COMPOUNDS WITH HIGH ENERGY POTENTIAL
A. Substrate Level Phosphrylation
B. Oxidative Phosphrylation
V. CHARACTERISTIC STRUCTURES
A. Enol Phosphate
B. Acid Anhydrides
C. Guanidinium Phosphates
D. Thiol Esters
VI. OXIDATIVE PHOSPHORYLATION
A. Electron Carriers (NAD and FAD: found in Mitochondria)
B. Participating Enzymes
CHAPTER 32: PORPHYRINS AND BILE PIGMENTS
INTRODUCTION
I. BIOMEDICAL IMPORTANCE
-Porphyrins and Iron are needed in the synthesis of Heme
-basic to understanding varied functions of Hemoproteins
A. Porphyrins -cyclic compounds formed by the linkage of four pyrrole rings through –HC=
Methenyl Bridges
-can form complexes with Metal Ions bound to the Nitrogen Atom of the Pyrrole Rings
-proteins that contain Heme (Hemoproteins) are widely distributed in nature
-ex) Iron Porphyrins: Heme
Magnesium-containing Porphyrin: Chlorophyll
Ferrochelatase
Protoporphyrin III (IX) Heme
Fe2++
A-Amino-B-Ketoadipate
(ALA Synthase)
Hydroxymethylbilane
Protoporphyrinogen III
Protoporphyrin III
(Ferrochelatase)
HEME
I. SUCCINYL – CoA + GLYCINE
-reaction occurs in the Mitochondria
-starting materials to synthesize Heme
-Succinyl CoA is derived from Citric Acid in the Mitochondria
V. SYNTHESIS OF UROPORPHYRINOGEN
A. Uroporphyrinogen I -is synthesized when HMB Cyclizes spontaneously
-auto-oxidized by light to its respective Porphyrins
A. Coproporphyrinogen III -enters the Mitochondria and converted into Protoporphyrinogen III
and then to Protoporphyrin III
B. Spectrophotometry
-used to test for Porphyrins and Precursors
-Coproporphyrins and Uroporphyrins are of clinical interest because they are excreted in high
amounts in the Prophyrias
Mutations in DNA
Abnormalities of the
Enzymes of Heme Synthesis
Photosensitivity
**Heme Oxygenase -complex enzyme system which carries out the catabolism of Heme
**Biliverdin Reductase -soluble enzyme which reduces Methenyl Bridge between Pyrroles to produce the
Bilirubin (yellow pigment)
-1 g of hemoglobin yields 35mg of Bilirubin
Bilirubin in Albumin
BLOOD
A. UPTAKE
Bilirubin
B. CONJUGATION HEPATOCYTE
Bilirubin Diglucuronide
C. SECRETION
BILE DUCTULE
Bilirubin Diglucuronide
A. Uptake of Bilirubin by the Liver
-in the liver, Bilirubin is removed from Albumin and taken up at sinusoidal surface of
hepatocytes by a carrier-mediated saturable system (Facilitated Transport System)
-once it enters Hepatocytes, it can bind to certain Cystolic Proteins
C. Secretion of Bilirubin
-secretion occurs as an active transport mechanism (Rate Limiting for the entire process of
Hepatic Bilirubin Metabolism)
-protein involved in secretion is MRP-2 (Multidrug Resistance-Like Protein 2) or MOAT
**Causes: 1. Increased production of Bilirubin that the normal liver can excrete (Hemolisis)
2. Failure of a damaged liver to excrete Bilirubin produced in normal amounts
E. Toxic Hyperbilirubinemia -due to hepatic parenchymal cell damage, which impairs conjugation
V. SUMMARY OF BIOSYNTHESIS:
• Hemoproteins (Hemoglobin and Cytochromes) contain heme (Heme is an iron-porphyrin compound in which
4 pyrrole rings are joined by Methenyl Bridges
• Biosynthesis of the Heme ring occurs in the Mitochondria and Cytosol via eight Enzymatic Steps
• Porphyrias result from abnormalities in heme biosynthesis (RBC and Liver Cell are the major sites of
Metabolic Expression of Porphyrias
• Catabolism of heme ring is inflated by the enzyme heme oxygenase, producing a linear tetrapyrrole
• Biliverdin is an early product of catabolism and on reduction yields Bilirubin
• In the Liver, Bilirubin is made water soluble by conjugation with 2 molecules of Glucuronic Acid and is
secreted in the Bile
• Jaundice is due to elevation of the level of Bilirubin in the Blood
HEMOGLOBIN AND ITS ROLE IN GAS TRANSPORT
I. BIOMEDICAL IMPORTANCE
-Myoglobin and Hemoglobin maintain a supply of Oxygen essential for Oxidative Metabolism
-illustrate both protein structure – function relationships and the molecular basis of genetic diseases
such as sickle cell disease and the thalassemias
II. HEMOGLOBIN
-effective carrier because oxygen is not soluble enough in blood plasma
-control oxygen delivery (since oxygen affinity is responsive to changing physiological conditions)
-Histidine is the Amino Acid Residue in Hemoglbin formation in Oxygen binding
-contains a Heme: cyclic tetrapyrrole with four molecules of Pyrrole and an atom of Fe2+ at the center
A. Cooperative Binding
-permits Hemoglobin to maximize both the quantity of Oxygen loaded at the PO2 of the lungs
and the quantity of Oxygen released at the PO2 of the Peripheral Tissues
-binding of First Oxygen to the deoxyHb molecule shifts the Heme ring and this motion is
transmitted to the Proximal Histidine
-this result to further conformational changes of the other subunits of hemogllbin
-critically important to Aerobic Life
**BPG or 2,3-Biphosphoglycerate
-the higher concentration, the more efficient in the delivery of Oxygen
-the concentration in the Red Blood Cell rise during tissue Hypoxia
-its high levels enhance formation of Deoxyhemoglobin at partial pressure of Oxygen
so that Hemoglobin then delivers more of its Oxygen to tissues
**BPG Mechanism will not compensate for tissue hypoxia when the Partial Pressure of Oxygen
in the lungs fall too low
!At low pressure of Oxygen in Lungs (cancer)
!Cannot be compensated if Pressure is too low
1. Carbamates -change the charge on amino terminals from (+) to (-), favoring salt bond
formation between the A and B chains
-account for about 15% of the Carbon Dioxide in the Venous Blood
-Carbon Dioxide attached to hemoglobin
3. Dissolved CO2
" CO2 generated in Peripheral Tissues combines with water to form Carbonic Acid
which dissociates into Protons and Bicarbonate ions
" Deoxyhemoglobin binds protons and deliver them to the lungs
" In the Lungs, the uptake of oxygen by Hemoglobin releases protons that combine
with Bicarbonate Ion, forming Carbonic Acid
" Carbonic Acid when dehydrated by Carbonic Anhydrase becomes Carbon Dioxide
and is then exhaled
E. Protons Arise from Rupture of Salt Bonds when Oxygen Binds to T-State Hemoglobin
-conversion to oxygenated R-State break salt bridges involving B-Chain residue at His146
-protons responsible for the Bohr Effect arise from rupture of salt bridges
-the dissociation of Protons from His 146 drives conversion of Bicarbonate to Carbonic Acid
-upon release of Oxygen, the T-Structure and its salt bridges reform
V. PHYSICAL FACTORS THAT AFFECT OXYGEN BINDING
B. High Levels of BPG -the higher concentration, the more efficient in the delivery of Oxygen
-stabilizes the T-State; Right shift
-in Hypoxia: produce more BPG and increased release of oxygen into tissues
!elevated BPG lowers the affinity of Hb Affinity for Oxygen (decreases P50) which enhances
the release of Oxygen at tissues
VI. SUMMARY OF OXYGEN – CARBON DIOXIDE BINDING
A. In the Lungs
**Conditions in the Lungs:
-high Pressure for Oxygen (around 100mmHg)
-high pH, low [H+]
-Hemoglobin has High Affinity for Oxygen
carbonic andhydrase
HCO3- + H+ H2CO3 CO2 + H20
(carbonic acid)
" The pressure for oxygen in lungs is high, which increases the Hemoglobin affinity for
oxygen. Oxygen binds to the Hemoglobin which releases the Protons (H+) and drives the
exhalation of CO2
" Increase in Pressure of Oxygen promotes proton release which in turn binds with
Bicarbonate to produce Carbon Dioxide
" Blood coming from heart is oxygenated and it unloads the oxygen in the tissues
" Deoxyhemoglobin binds one proton for every two Oxygens released, contributing to the
buffering capacity of blood
" Lower pH in tissues (aided by Carbamation) stabilizes the T-State and enhances the delivery
of Oxygen
" In the tissues, affinity for oxygen is low which caused the release of Oxygen
VII. MUTANT HUMAN HEMOGLOBINS (not bind with Oxygen)
C. Hemoglobin-S -the non-polar amino acid valine has replaced the polar surface residue GluG of
the B-Subunit
-Hydrophobic “Sticky Patch” on the surface of B-Subunit of both oxyHbs and
deoxyHbs
-distort the erythrocytes into sickle shape rendering it vulnerable to Lysis in the
Splenic Sinusoides (low PO2 exacerbates the tendency to polymerize)
A. Buffering -Hb’s buffering mechanism at ionizable groups with pK values close to the
Intraerythrocute pH
-more of Hb’s buffering ability is provided by the Imidazole side chains of the
Histidine Residues (38 Histidine per Hb Tetramer)
B. Isohydric Mechanism -Hb becomes a stronger acid and releases H+ when it becomes oxygenated
(Bohr Effect)
-in the capillaries, where O2 is released from Oxyhemoglobin:
HbO2 + H+ HHb + O2
A. Cyanosis -patient’s skin or mucous membrane appears gray or (in severe cases) purple
-due to an abnormally high concentration of Deoxyhemoglobin below surface
which is responsible for color
-most commonly caused by diseases of the Cardiac or Pulmonary Systems,
resulting in inadequate Oxygenation of the blood
-in someone with low Hb, it cannot lead to Cyanosis because there is no Hb to
begin with (there will be Anemia)
INTRODUCTION
I. METABOLISM (Catabolism and Anabolism make-up Metabolism)
**Not all energy are trapped by the body (only 60% are trapped)
**Energy not trapped escapes as Heat (form of energy which cannot be utilized by the Body)
B. Law of Entropy
-if we leave a system on its own, it would always proceed to a state of equilibrium
-state of equilibrium: reactant forming a product; product forming a reactant
-disadvantage of equilibrium: No work is performed (nothing accomplished) Wasting energy!
A. Gibb’s Equation
D. !G0 and Keq (when concentration of product and reactant are known, !G0 can be obtained)
A+B C+D
Keq = ( C ) ( D )
(A) (B)
**Coupled Reactions -product of the first reaction must be the substrate of the second reaction
-Endergonic Reaction must be coupled to an Exergonic Reaction to proceed
-amount of energy endergonic needed must be given by exergonic reaction
-amount of energy liberated by exergonic reaction must be enough to drive
the products of endergonic
-ex) Glycolysis
**Glycolysis
+ Pi
1. Glucose Glucose – 6 – Phosphate !G = + 3.3 Kcal / mol (Endergonic)
+ H2O
2. ATP ADP + Pi !G = - 7.3 Kcal / mol (Exergonic)
B. Oxidative Phosphrylation
-synthesis by Oxidation of the Substrate and the Oxidation liberated the energy
-the Energy is trapped and used in the Synthesis of ATP
V. CHARACTERISTIC STRUCTURES
-used by the body in Substrate Level Phosphorylation
A. Enol Phosphates
o Phosphoenolpyruvic Acid
B. Acid Anhydrides
o 1,3 Biphosphoglyceric Acid (high energy bond: 1)
o ATP, GTP, TTP, CTP
C. Guanidinium Phosphates
o Creatine Phosphate (in muscles for more ATP)
D. Thiol Esters
o Acetyl Coenzyme A (from Phantothenic Acid: Vitamin B)
**Superoxide Anion Oxidase -specific enzyme that protects tissues from oxygen toxicity caused
by free radicals
VI. OXIDATIVE PHOSPHORYLATION
-ATP Synthesis Via the Respiratory Chain (chain involved in cellular respiration)
-ATP Synthesis thru oxidation of substrates
-Oxidation of substrates releases electrons
-Electrons are carried by NADH and FADH2 (carriers of Electrons)
1. NAD -accepts a Hydride Ion (Hydrogen Atom with one extra electron: H+ +)
-carrier of Electrons in Oxidation
-forms NADH + H+ when it accepts two electrons and one proton (the other
Proton is released in the Medium)
BIOLOGIC OXIDATION
INTRODUCTION
• Source of energy for the body is from the Oxidation of Foods
• All molecules can be degraded into a common compound: Acetyl-CoA
• Acetyl-CoA is a high energy compound (2-Carbon High Energy Component)
• Acetyl-CoA enter TCA and the 2-Carbons are liberated as CO2
• The main purpose is to Yield energy
• Trapping of energy = Trapping of Electrons and are taken up by Electron Acceptors (NAD / FAD)
• Electrons are delivered into an Electron Transport System (ETC) / Respiratory Chain
• Transfer of Electrons from NADH & FADH2 from one member to another until it reaches the last
member: Cytochrome Oxidase
• When electrons are delivered to Oxygen, Water is formed
• When Electrons are moving, Reduction Potential is produced and free energy is released to form the
Phosphodiester Bond to form ADP + Pi to ATP
Pyruvate
Acetyl-CoA
O
e- ! ETC ! !
H 2O
ADP + Pi ATP
PARTICIPATING ENZYMES IN BIOLOGIC OXIDATION
-enzymes involved in Oxidation and Reduction are called Oxidoreductases
-classified into four groups: Oxidases, Dehydrogenases, Hydroperoxidses, Oxygenases
I. OXIDASES
-can use oxygen as a Hydrogen Acceptor
-catalyze removal of Hydrogen from a substrate using Oxygen as an acceptor
-form water of Hydrogen Peroxide as a product
A. Cytochrome Oxidase -hemoprotein distributed in many tissues having the typical heme Prosthetic
group (such as in Hemoglobin, Myoglobin, Cytochrome)
-terminal component of Respiratory Carriers in mitochondria
-transfers electrons from oxidation of substrates by dehydrogenases to the
final acceptor: Oxygen
-poisoned by Carbon Monoxide, Cyanide, Hydrogen Sulfide
-also known as Cytochrome aa3
-two atoms of Copper are present associated with a heme unit
B. Flavoprotein Enzymes -contain Flavin Mononucleotide (FMN) / Flavin Adenine Dinucleotide (FAD)
-ex) L-Amino Acid Oxidase (FMN linked enzyme)
Xanthine Oxidase (conversion of purine bases)
Aldehyde Dehydrogenase (FAD linked enzyme)
II. DEHYDROGENASES
-cannot use oxygen as a Hydrogen Acceptor
-perform two main functions:
1. Transfer of Hydrogen from one substrate to another in a coupled redox reaction
2. Components in the Respiratory Chain of Electron Transport from substrate to Oxygen
III. HYDROPEROXIDASES
-use hydrogen peroxide or an organic peroxide as substrate and protect body against harmful peroxides
-two types of enzymes found in both animals and plants: Peroxidases and Catalase
IV. OXYGENASES
-catalyze direct transfer and incorporation of oxygen into a substrate molecule in two steps:
1. Oxygen is bound to enzyme at the active site
2. Bound oxygen is reduced or transferred to the substrate
-can be divided into: Dioxygenase and Monooxygenases
!Oxidative Phosphorylation
!The Electron Transport Chain
!Importance
I. OXIDATIVE PHOSPHORYLATION
-oxidation of substrates, coupled by the synthesis of ATP
-if ATP Synthesis is not coupled to Oxidation of Substrates, the energy synthesized by oxidation is
liberated as Heat
-without oxidation, there will be no Phosphorylation (dependent on Oxidation)
-Oxidation is Independent on Phosphorylation, but Phosphorylation is dependent on Oxidation
-rate of Oxidation will increase if there is no Phosphorylation
e -s
Substrate H2 H2
(reductant: compound w/ 2e-)
Oxidized Substrate
A. The Mitochondria
-powerhouse of the cell: site of ATP synthesis
-has an outer membrane, inner membrane, matrix within
-there is also ATP production in cytoplasm (anaerobic: not so efficient)
-Cristae increases surface area to have much enzymes to synthesize ATP
**Thermogenin -found in the membrane and instead of the Proton passing from
intramembranous space to the matrix, some protons are attracted to
the Thermogenin
-Protons which enter Thermogenin will not produce a Proton Motive Force
-Proton Motive Force + ATPsynthase = ATP synthesis
-without either the Force or ATPsynthase = NO ATP Synthesis