Valerie O.

Yazbeck
Student ID: 40170951
EXCI 258 Group 2, October 18th

Table 1:
Dilution
Table
Dilution Volume Volume [NADH] Absorbance Absorbance Average
Factor Stock H2O #1 #2 Absorbance
10 300 2700 5 0.128 0.128 0.128
5 600 2400 10 0.259 0.257 0.258
2.5 1200 1800 20 0.511 0.492 0.502
2 1500 1500 25 0.641 0.644 0.643
unknown 15 0.381

Graph 1:

Concentration of NADH vs the Average Absorbance

0.5

0.4

0.3 Series1
Linear (Series1)
0.2

0.1

0
0 5 10 15 20 25 30
[NADH] (mM)

Short Answer Questions:

1. What is NAD/ NADH and where is it found in muscle fibers?

• NAD+, or nicotinamide adenine dinucleotide, is a coenzyme found that is found in every single
living cell. NAD exists in two forms, NAD+ and NADH and is a central metabolic
coenzyme/cosubstrate involved in cellular energy metabolism and energy production. NAD+ can
be readily be reduced by two electron equivalents which gives us the NADH form, which is the
 minority species to NAD+ under most physiologic conditions. Additionally, NAD/NADH is
found in the mitochondria within muscle fibers.

2. What is its role? What does it do?

• NAD+ plays an important role in not only oxidation-reduction reactions in cells but also as a
signaling molecule. NAD+ is an electron carrier which will pick up electrons during the course
of cellular respiration. When NAD+ picks up an electron, it becomes reduced, and becomes
NADH. NADH carries electrons all the way to the Electron Transport Chain, where it will then
drop off the electrons. This will allow for the powering of protons to be pumped across a
concentration gradient, which then allows ATP Synthase to churn, producing ATP in the end.
Moreover, NAD+ plays a key role in mitochondrial function via participation in pyruvate
dehydrogenase, tricarboxylic acid cycle, and oxidative phosphorylation chemistries. It also acts
as a substrate for deacylases SIRT3, SIRT4, and SIRT5, which modify protein posttranslational
modifications on lysine within the mitochondrial compartment. Additionally, NADH is a
coenzyme in making ATP. It exists in two forms in the cell: NAD+ and NADH where the first
form, NAD+, is called the oxidized form. When a molecule is in an oxidized state, it means it
can accept electrons from another molecule. After it gets the electrons, it has a negative charge,
so it also picks up a hydrogen atom from the surrounding environment, since hydrogen atoms are
positively charged. Now, we have the reduced form, or NADH. The molecule acts as a shuttle
for electrons during cellular respiration. During chemical reactions, the NAD+ picks up an
electron from glucose, at which point it becomes NADH. Then NADH, along with another
molecule flavin adenine dinucleotide (FADH2) will ultimately transport the electrons to the
mitochondria, where the cell can harvest energy stored in the electrons.

3. What is the functional difference between FAD and NAD?

• FAD, which stands for flavin adenine dinucleotide, and NAD which is nicotinamide
adenine dinucleotide are both electron carriers which have many roles to perform.
Nicotinamide adenine dinucleotide is a coenzyme that is found in all living beings. In
regards to NAD, the nucleotides are joined together by phosphate groups. While, FAD is
considered a redox cofactor that is related to many metabolic reactions. FAD also exists
in two redox states. One of the main differences that can be seen between FAD and NAD
is in the difference in how they each accept hydrogen atoms. FAD can accommodate two
hydrogens whereas NAD accepts just one hydrogen. In NAD, a single hydrogen and an
electron pair is transferred, and the second hydrogen is freed, hence why the reduced
FAD becomes FADH2, and the reduced NAD becomes NADH + H+. NAD gets reduced
to NADH during the metabolic processes, Kreb’s cycle and glycolysis. It feeds into the
electron transport chain at Complex 1 and also gives 3 ATP for every NADH. FAD is
normally reduced to FADH2 in the Krebs cycle and it feeds into the electron transport
chain at Complex 11 and also gives 2 ATP for every FADH2. During electron transfer, the
FAD and NAD reduce cytochromes differently. While NADH+H+ reduces Cytochrome I,
FADH2 is known to reduce Cytochrome II. (S, P. (2011, July 28). Difference Between
NAD and FAD. Difference Between Similar Terms and Objects.
http://www.differencebetween.net/science/difference-between-nad-and-fad/.)
References:

• . (S, P. (2011, July 28). Difference Between NAD and FAD. Difference Between Similar
Terms and Objects. http://www.differencebetween.net/science/difference-between-
nad-and-fad/.)