Seminario Biologia molecular Estefania Blandon Hurtado UPB

1. Estefanía Blandón Hurtado
Medicine III semester
UPB 2023-2

2. It is an extremely important cell organelle in the production of
energy.
Mitochondria occupy 15% to 20% of the volume of an average
mammalian liver cell and contain over a thousand different
proteins.
Some features:
• Generates energy -> Adenosine triphosphate (ATP)
• It is inherited from the mother.
• It has its own DNA
INTRODUCTION
Respiración aeróbica y la mitocondria. En: McGraw Hill Medical.

3. CATABOLISM
In the BCAA catabolization process is a-Keto acid
dehydrogenase (BCKD), it is an enzyme located in the
mitochondrial membrane that irreversibly catalyzes
decarboxylation. BCKD is regulated by the
phosphorylation-dephosphorylation mechanism
Reactions by which organic nutrients are broken down
into simple products, in order to extract chemical energy
from them and convert it into a useful form for the cell
del Ejercicio F. Aminoácidos de cadena ramificada en la salud y enfermedad: metabolismo, alteraciones en el plasma
sanguíneo, y como suplementos [Internet]. Fisiología del Ejercicio. 2018 [citado el 27 de julio de 2023].
BCAA BCKD -> Oxidation

4. GENERAL OBJETIVE
The mitochondrial pyruvate carrier (MPC) has emerged as a therapeutic target for treating insulin
resistance, type 2 diabetes, and nonalcoholic steatohepatitis (NASH). We evaluated whether MPC
inhibitors (MPCi) might correct impairments in branched chain amino acid (BCAA) catabolism, which are
predictive of developing diabetes and NASH
Mitochondrial%20pyruvate%20carrier%20inhibition%20initiates%20metabolic%20crosstalk%20to%20stimulate%20branched%20chain%20amino%20acid%20catabolism.pdf

5. Basis It is an in vitro study model made up of
cells that can grow and remain in suspension for
more than 24 hours under controlled conditions.
MATERIALS AND METHODS
CELL CULTURE EXPERIMENTS BCKA OXIDATION ASSAY
Basis Protein oxidation involves the initial
modification of amino acid side chain groups by
ROS and subsequent conversion to carbonyl
What for ? BCKA was oxidized to normalize
the protein concentration after having isolated
the cells with the treatment, after this oxidation it
was evidenced that the protein levels were
normalized
What for ?  Human hepatoma and also
mouse hepatocytes are cultured as control, with
and without treatment for protein determination.

6. Basis  It is a technique used in molecular
biology to identify specific proteins, it uses a
highly sensitive system for the detection of
proteins through the recognition between antigen
and antibody.
IMMUNOBLOTTING
mTOR PHOSPHO
ANTIBODY ARRAY
Basis  It is an ELISA-based antibody matrix for
the qualitative profiling of protein phosphorylation.
compare normal samples with treated or diseased
samples and identify candidate biomarkers. .
AllScience. Introducción a Western Blot [Internet]. AllScience. [citado el 22 de octubre de 2022]. Disponible
en: https://www.e-allscience.com/blogs/articulos/introduccion-a-western-blot
MTOR Phospho Antibody Array [Internet]. Full Moon BioSystems. 2014 [citado el 27 de julio de
2023].
What for? cell signaling
with protease and
phosphatase inhibitors to be
able to analyze the specific
proteins of non-
phosphorylated amino acids.
tyrosine serine Threonine
What for? Human hepatoma cells were treated
with and without treatment. mTOR samples were
used for observation of specific antibodies to
reveal phosphorylation levels and patterns in a
single assay.

7. Result

9. D
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10. Conclusions
Molecular biology has revolutionized medicine. The study of genes and proteins has led to a greater understanding of
inherited diseases, allowing the development of more precise genetic tests and treatments. In addition, molecular biology has
been crucial in the development of gene therapies and drugs specifically directed at molecular targets, as we can see in this
study.
The balance between intake and catabolism of branched-chain amino acids is the starting point for reducing the risk of
diabetes and non-alcoholic hepatitis, since, as the article mentions, “if BCKAs are not oxidized, BCAA accumulation
increases, generating a greater risk of obesity and DM, this is where molecular tests are essential for a good study on MPC
inhibition since it decreases hepatic glucose production
By reducing the amount of pyruvate entering the mitochondria, ATP production is reduced. This could activate a cellular
response that increases insulin sensitivity in cells. Increased insulin sensitivity allows for better uptake and utilization of
glucose by cells, which could help prevent insulin resistance and therefore the risk of type 2 diabetes, it is necessary that with
the study of molecular biology an approach is sought to compensate for the loss of ATP by inhibiting MPC, perhaps activating
other metabolic pathways