Lactate dehydrogenase is a
vital enzyme in the process of anaerobic cellular respiration. Anaerobic
cellular respiration is an important function in plants, animals, and bacteria
to produce ATP. Lactate dehydrogenase is found in almost all living cells to
serve as a catalyst for anaerobic cellular respiration.


         The objective of studying lactate dehydrogenase is to learn
about the structure, function, and importance. The goal is to become familiar
with the enzyme and the metabolic processes that it is involved with. Without
lactate dehydrogenase anaerobic cellular respiration would not occur. This
would substantially inhibit the production of ATP by the cell. Some bacteria
rely solely on anaerobic cellular respiration as the main source of ATP, and without
lactate dehydrogenase there would be no energy production in the cell.

Metabolic Pathway of
Lactate Dehydrogenase

respiration that does not require oxygen is defined as anaerobic cellular
respiration. A final acceptor is needed at the end of the electron transport
chain. The final acceptor in aerobic cellular respiration is oxygen, but in
anaerobic cellular respiration the final acceptor is a less-oxidizing compound.
Less energy is formed from each oxidized molecule since these molecules have a
smaller reduction potential than oxygen. Anaerobic cellular respiration is much
less efficient when compared to aerobic cellular respiration. Anaerobic
cellular respiration functions to produce lactate acid from pyruvate with no
oxygen present. Anaerobic cellular respiration is important for glycolysis. The
accumulation of pyruvate would slow down ATP production. Anaerobic cellular respiration
functions to regenerate NAD+ from NADH. In humans, as one exercises, glucose is
completely broken down which releases carbon atoms as carbon dioxide and
hydrogen molecules as water. This process requires substantial amounts of
oxygen. Energy production will stop at the end of glycolysis if the supply of
oxygen does not meet the demand for oxygen. Energy can still be produced when
the supply of oxygen does not meet the demand through anaerobic cellular
respiration. However, this process is less efficient and less ATP is produced.
Lactate dehydrogenase makes this process possible.

dehydrogenase is a key enzyme that is involved with anaerobic cellular respiration.
As stated above anaerobic cellular respiration is key in the regeneration of
NAD+ from NADH. Lactate dehydrogenase is the main enzyme involved with
converting NADH to NAD+. Lactate dehydrogenase converts lactate to pyruvic acid
and back to lactate as the conversion of NADH to NAD+ is occurring. During glycolysis,
the hydrogen atom from glucose is put on NAD+ and forms NADH. These hydrogen
atoms are transferred to oxygen to form water when oxygen is available,
however, when oxygen is unavailable, the NADH will build up and there is not
enough NAD+ to continue producing ATP using glycolysis. Lactate dehydrogenase
combines pyruvate and the built up NADH to form lactic acid and NAD+. This NAD+
formed can then be used to complete another cycle of glycolysis, thus producing
more ATP. This process quickly creates more energy.

The Gene Ontology Terms

The biological processes for
lactate dehydrogenase according to gene ontology are vast. The biological
processes of lactate dehydrogenase include: response to hypoxia, carbohydrate
metabolic process, lactate metabolic process, pyruvate metabolic process,
glycolytic metabolic process, response to nutrient, response to glucose,
response to organic cyclic compound, NAD metabolic process, carboxylic acid
metabolic process, response to drug, response to hydrogen peroxide, positive
regulation of apoptotic process, response to estrogen, post-embryonic animal
organ development, response to cAMP, and oxidation-reduction process. Lactate
dehydrogenase can be found throughout the cell. According to gene ontology
lactate dehydrogenase is found in the following locations in the cell: nucleus,
cytoplasm, cytosol, membrane, and integral component of membrane. It has been
seen that LDH has many molecular functions. Some of the molecular functions
are: catalytic activity, lactate dehydrogenase activity, L-lactate
dehydrogenase activity, protein binding, oxidoreductase activity, acting on the
CH-OH groups of donors, NAD or NADP as an acceptor, kinase binding, identical
protein binding, cadherin binding, and NAD binding.

History of Isolation

         Human LDH-X was isolated from frozen samples of semen using
affinity chromatography. When NAD+ is present the LDH-X does not bind to
AMP-Sepharose. The other lactate dehydrogenase isoenzymes will bind to
AMP-Sepharose. This is the key point in isolating LDH-X versus the other

frozen semen samples were thawed and centrifuged at 30,000 g and four degrees
Celsius for 20 minutes. Approximately 500 milliliters of the seminal fluid were
separated by ammonium sulfate. The precipitate that was formed was dialyzed
against a sodium phosphate buffer. The sodium phosphate buffer had a pH of 6.8.
This same buffer was used for all of the chromatography steps. The temperature
was kept at 4 degrees Celsius for the entire procedure. In the presence of
NADH, lactate dehydrogenase isoenzymes will bind to the column and are then
eluted by the buffer. In the presence of buffer only, lactate dehydrogenase
isoenzymes will also bind to AMP-Sepharose. It was found that if equal volumes
of seminal fluid and buffer containing NADH were mixed immediately before
loading it into the column, enough NADH was still present to allow complete
binding of lactate dehydrogenase to the column. AMP-Sepharose was used to
separate LDH-X from the other LDH isoenzymes since LDH-X does not bind to

Characteristics of the

lactate dehydrogenase protein contains a disordered portion of approximately 50
residues. This disordered region has discontinuous electron density. The
lactate dehydrogenase protein model contains: residues 9-328, 375-567, an
acetate molecule, a FAD molecule, and approximately 200 water molecules for
each monomer. The two monomers are basically identical.

The lactate dehydrogenase
protein is made up of three discontinuous domains: the FAD-binding domain
(residues 1–268 and
520–571), the cap domain (residues 269–310, 388–425, and 450–519), and the
membrane-binding domain (residues 311–387 and 426–449, residues 329 –376 are in the
disordered region). The FAD-binding domain contains two alpha + beta
subdomains. The first subdomain is made up of three antiparallel beta beta
strands surrounded by five alpha helices and is packed closely to the second
domain. The second subdomain is made up of five parallel beta strands
surrounded by four alpha helices. The cap domain is composed of a large seven
stranded antiparallel beta sheet that is surrounded on both sides by alpha
helices. Four alpha helices make up the membrane binding domain. The largest
difference between these structures is in the membrane-bounding domain.

dehydrogenase is considered to be a part of the FAD-containing family. The main
difference between LDH and other members of the FAD-containing family is the
membrane binding domain. In other proteins that are classified in the
FAD-containing family, the membrane binding domain is either not present or
much different. An electropositive surface with five Lys residues and six Arg
residues make up the membrane binding domain of lactate dehydrogenase. The
residues that make up the membrane binding domain are expected to interact with
the negatively charged phospholipid head groups of the membrane. Rather than
binding to the membrane with hydrophobic forces, lactate dehydrogenase binds to
the membrane with electrostatic forces. Some other members of the
FAD-containing protein family are: vanillyl-alcohol oxidase, p-cresol
methylhydroxylase (PCMH), and UDP-N-acetylenolpyruvyglucosamine (MurB). The
proteins in this family can be found in both eukaryotes and eubacteria.

Characteristics of the
Gene for Lactate Dehydrogenase

The LDHA gene in humans is
located on chromosome 11p15.4. Chromosome 11 is approximately 135 million base
pairs and accounts for around 4-4.5 percent of DNA in the cells. Chromosome 11
contains approximately 1,300-1,400 genes that give instructions for
synthesizing proteins. These proteins have a wide array of tasks in the body.
The LDHB gene is located on chromosome12p12.2-p12.1. Chromosome 12 is made up
of almost 134 million base pairs and accounts for around 4-4.5 percent of the
DNA in cells. Chromosome 12 contains approximately 1,100-1,200 genes that
provide instructions for synthesizing proteins. These proteins also also have a
wide array of tasks in the body. The LDHC gene is only expressed in the testes
and can be found on chromosome 11p15.5-p15.3. The human genome also has several
non-transcribed LDHA pseudogenes. M subunit mutations have been observed to be
disease causing, H subunit mutations have not been linked to a certain disease
causing trait. LDHA mutations have been linked to cause exertional
myoglobinuria and Fanconi-Bickel Syndrome.

There are four genes for
lactate dehydrogenase: LDHA, LDHB, LDHC, and LDHD. LDHA, LDHB, and LDHC are the
L-isomers. LDHD is a D-isomer. The L-isomers use and produce L-lactate.
L-lactate is the major enantiomer found in vertebrates. LDHA is called the M
subunit and is mostly found in skeletal muscle. LDHB is called the H subunit
and is mostly found in the heart. Five isoenzymes can be formed from the M and
H subunits of LDH. The isoenzymes are: LDH-1 (4H), LDH-2 (3H, 1M), LDH-3 (2H,
2M), LDH-4 (1H, 3M), and LDH-5 (5M). LDH-1 and LDH-5 have the same active site
region. These isoenzymes are similar in function but have a different
distribution throughout tissues.

Regulation of the
Enzyme at Transcriptional and Enzymatic Levels

LDHA promoter region is well known to contain the consensus sequences for, and
be regulated by, major transcription factors: hypoxia-inducible factor 1 (HIF1)
and c-Myc. Forkhead box protein M1 (FOXM1) and Kruppel-like factor 4 (KLF4) are
identified as transcriptional regulators of LDHA. The regulation of LDHA is
very complex. Complete understanding of how LDHA is regulated is far from being
achieved. It has also been found that LDHA transcription is influenced by other
factors such as: lactate, cyclic adenosine monophosphate (cAMP), estrogen,
ErB2, and heat shock factor. It is highly likely that transcriptional
regulation of LDHA is influenced by many other unknown factors. Like many other
known enzymes, the post-transcriptional activity of LDHA is regulated by the
phosphorylation and acetylation of amino acid residues. PGC-1? regulates
lactate dehydrogenase at a transcriptional level. By decreasing LDHA mRNA
transcription and the enzymatic activity of pyruvate to lactate conversion,
PGC-1? regulates lactate dehydrogenase.

the enzymatic level, LDH is regulated by the relative concentrations of its
substrates. When there is major muscular output this creates an increase of
substrates available for the lactate dehydrogenase reaction, causing lactate
dehydrogenase to become more active. The demand for ATP increases when the
muscles are forced to produce a large amount of power. This demand causes a
buildup of free Pi, AMP, and ADP. The glycolytic flux that occurs due to this
buildup makes it difficult for certain shuttle enzymes to metabolize pyruvate. In
response to increased levels of pyruvate and NADH, the flux through lactate
dehydrogenase increases to metabolize pyruvate into lactate.


         There are many more processes lactate dehydrogenase is
believed to be involved with. This enzyme will continue to be further studied
in hopes of being targeted for certain disorders. Recent research has shown
lactate dehydrogenase to be a therapeutic target for certain types of cancers.
This gives hope that lactate dehydrogenase could be a potential target for the
treatment of cancers and cancer associated disorders. There are vast
pharmacological applications to be considered from this research. It can be
seen how important lactate dehydrogenase is in the cell. 


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