Sikandar

Ali
ROLL NO.105

1-Illustrate oxidative and non-oxidative reactions of HMP pathway.

2-Give the significance of HMP shunt.

 The hexose monophosphate shunt, also known as the pentose phosphate pathway, is a unique
pathway used to create products essential in the body for many reasons.
The HMP shunt is an alternative pathway to glycolysis and is used to produce ribose-5-phosphate
and nicotinamide adenine dinucleotide phosphate (NADPH).

3-Enumerate the uses of NADPH.

Free radicals scavange.

Fatty acid synthesis

Steroid synthesis

Nucleotide synthesis

Bile synthesis

4-Describe the role of NADPH with reference to cytochrome P450 enzyme system.

Monooxygenases (mixed-function oxidases) incorporate one atom from molecular oxygen into a
substrate , with the other atom being reduced to water. In the cytochrome P450 monooxygenase system, NADPH
provides the reducing equivalents required by this series of reactions. This system performs different functions in
two separate locations in cells. The overall reaction catalyzed by a cytochrome P450 enzyme is:
R-H + O2 + NADPH + H+ → R-OH + H2O + NADP+
where R may be a steroid, drug, or other chemical.

5-Describe and illustrate the oxygen dependent bactericidal mechanisms.

Glutathione (GSH) is essential for red cell integrity and is maintained in its reduced (functional) form
by nicotinamide adenine dinucleotide phosphate (NADPH)-dependent glutathione reductase. The NADPH is
generated by the oxidative portion of the pentose phosphate pathway. Individuals with a deficiency of the initiating
and regulated enzyme of this pathway, glucose 6-phosphate dehydrogenase (G6PD), have a decreased ability to
generate NADPH and, therefore, a decreased ability to keep GSH functional. When treated with an oxidant drug
such as primaquine, some patients with G6PD deficiency develop a hemolytic anemia. Primaquine does not affect
glucose 6-phosphate levels.

6-What are factors precipitating symptoms in G6PD deficiency and why?

1. Oxidant drugs: Commonly used drugs that produce hemolytic anemia in patients with G6PD
deficiency are best remembered from the mnemonic AAA: antibiotics (for example, sulfamethoxazole and
chloramphenicol), antimalarials (for example, primaquine but not chloroquine or quinine), and antipyretics (for
example, acetanilide but not acetaminophen).
 2. Favism: Some forms of G6PD deficiency, for example the Mediterranean variant, are particularly
susceptible to the hemolytic effect of the fava (broad) bean, a dietary staple in the Mediterranean region. Favism, the
hemolytic effect of ingesting fava beans, is not observed in all individuals with G6PD deficiency, but all patients
with favism have G6PD deficiency.

3. Infection: Infection is the most common precipitating factor of hemolysis in G6PD deficiency. The
inflammatory response to infection results in the generation of free radicals in macrophages, which can diffuse into
the RBC and cause oxidative damage.

7-What is the role of transaldolase and transketolase in the metabolism of glucose?

Transketolase (which transfers two-carbon units in a thiamine pyrophosphate [TPP]-requiring reaction)and

transaldolase (which transfers three-carbon units) convert the ribulose 5-phosphate produced as an end product of
the oxidative reactions to glyceraldehyde 3-phosphate and fructose 6-phosphate, which are glycolytic intermediates.

8-A patient after taking antimalarial drugs developed severe intravascular hemolysis.
Name the enzyme deficiency, how hemolysis occurred in RBC?
Glutathione (GSH) is essential for red cell integrity and is maintained in its reduced (functional) form by
nicotinamide adenine dinucleotide phosphate (NADPH)-dependent glutathione reductase. The NADPH is generated by the
oxidative portion of the pentose phosphate pathway. Individuals with a deficiency of the initiating and regulated enzyme of
this pathway, glucose 6-phosphate dehydrogenase (G6PD), have a decreased ability to generate NADPH and, therefore, a
decreased ability to keep GSH functional. When treated with an oxidant drug such as primaquine, some patients with G6PD
deficiency develop a hemolytic anemia. Primaquine does not affect glucose 6-phosphate levels.
Mutation in gene for G6PD, decrease enzyme activity, decrease production of NADPH, decrease reduced
glutathione, increase reactive oxygen intermediates, increase damage to RBC membrane, increase hemolysis.

9-Describe different types of G6PD deficiency diseases. What type of mutations cause
results in deficiency of the enzyme?
Variant enzymes may show decreased catalytic activity, decreased stability, or an alteration of binding affinity for
NADP +, NADPH, or glucose 6-phosphate. The severity of the disease usually correlates with the amount of residual enzyme
activity in the patient’s RBC. For example, variants can be classified as shown in. G6PD A– is the prototype of the moderate
(Class III) form of the disease. The RBCs contain an unstable but kinetically normal G6PD, with most of the enzyme activity
present in the reticulocytes and younger RBCs . The oldest RBCs, therefore, have the lowest level of enzyme activity and are
preferentially removed in a hemolytic episode. G6PD Mediterranean is the prototype of a more severe (Class II) deficiency in
which the enzyme has decreased stability resulting in decreased enzymic activity. Class I mutations (rare) are the most severe
and are associated with chronic nonspherocytic hemolytic anemia, which occurs even in the absence of oxidative stress.
10-How nitric oxide (NO) is synthesized in our body? Describe its role in vasodilation

and smooth muscle relaxation.

Arginine, O2, and NADPH are substrates for cytosolic NO synthase . FMN, FAD, heme, and tetrahydrobiopterin are
coenzymes, and NO and citrulline are products of the reaction. Three NOS, each the product of a different gene, have been
identified. Two are constitutive, Ca2+calmodulindependent enzymes. They are found primarily in endothelium and neural
tissue and constantly produce very low levels of NO for vasodilation and neurotransmission. An inducible, Ca2+-independent
enzyme can be expressed in many cells, including macrophages and neutrophils, as an early defense against pathogens. The
specific inducers for iNOS vary with cell type, and include proinflammatory cytokines, such as tumor necrosis factor-α and
interferon-γ, and bacterial endotoxins such as lipopolysaccharide. These compounds promote synthesis of iNOS, which can
result in large amounts of NO being produced over hours or even days.
Actions of nitric oxide on vascular endothelium: NO is an important mediator in the control of vascular smooth
muscle tone. NO is synthesized by eNOS in endothelial cells and diffuses to vascular smooth muscle, where it activates the
cytosolic form of guanylate cyclase to form cyclic cGMP. The resultant rise in cGMP causes activation of protein kinase G,
which phosphorylates Ca2+ channels, causing decreased entry of Ca2+ into smooth muscle cells. This decreases the calcium–
calmodulin activation of myosin light-chain kinase, thereby decreasing smooth muscle contraction and favoring relaxation.
Vasodilator nitrates, such as nitroglycerin, are metabolized to NO, which causes relaxation of vascular smooth muscle and,
therefore, lowers blood pressure. Thus, NO can be envisioned as an endogenous nitrovasodilator.