Pyridoxal Phosphate, PLP

Transamination only 1 of many a.a. transformations it

can catalyze at alpha carbon
Decarboxylations, deaminations, racemizations,
aldol cleavages
Can also catalyze elimination/replacement reactions
at -Cs
All reactions similar--go via Schiff base w/a.a.
substrate, protonated PLP acts as electron sink to
stabilize neg. charged intermediates (ring N attracts es
from a.a.), and Schiff base hydrolyzed giving desired
product
Bond cleaved determined by active site configuration

Amine groups removed are transferred to -ketoglutarate to

form glutamate in the cytosol, then are eventually removed by
oxidative deamination after glutamate is moved to the matrix

Deamination contd
Ammonia formed in tissues is transported to the
liver or kidneys for removal
Ammonia is converted into a non-toxic compound
for transport in the blood, glutamine or alanine
Ammonia is coupled to glutamate though a two
step, glutamine synthetase catalyzed reaction

Ammonia in tissue can be

transported to liver using
glutamine

 Alanine is a another non-toxic carrier of ammonia

that transports ammonia to the liver
In the process pyruvate is also transported and is
used for the synthesis of more glucose to replenish
muscle glucose and glucose stores as part of the
glucose-alanine cycle
Glutamate can also transfer its amino group to
pyruvate to form alanine

What happens to NH4+ in the liver?

Ammonotelic animals--secrete to water
Most aquatic vertebrates

Uricotelic animals--uric acid

Birds, reptiles

Ureotelic animals--urea
Many terrestrial vertebrates (also sharks)
Urea cycle

Urea Cycle--Urea Synthesis

Hans Krebs & Kurt Henseleit (1932)
Urea synthesis in liver main way body clears NH4+
2 N atoms incorporated, C comes from CO2
One N comes from transamination, other from NH4+
Ornithine (non-conventional amino acid) carrier of N & C
atoms, citrulline 2nd non-conventional amino acid
Overall reaction
CO2 + NH4+ + 3 ATP + 2H2O + Asp urea + 2ADP +
2Pi + AMP + PPi + fumarate, and PPi hydrolyzed
4 ATP equivalents used to make 1 urea
Production of fumarate links urea cycle with Krebs cycle
fumarate --> OOA --> Asp, GNG, Krebs cycle or converted to Pyr

One copy of ammonia enters

the urea cycle in its first step
as carbamoyl phosphate;
carbamoyl phosphate requires
two ATPs to be produced.
First activates bicarbonate;
second phosphorylates
carbamate.

1. ornithine
transcarbamolase

2. argininosuccinate
synthetase
4. arginase

3. argininosuccinase

Some Initial Mechanisms in the Urea Cycle

The Primary Regulation of the Urea Cycle is the

Rate of Formation of Carbamoyl Phosphate which is
Regulated by N-acetylglutamate

Interaction of Urea and Citric Acid Cycles

Fate of Amino Acids

Ten of the amino acids are eventually converted to
acetyl CoA (ala,cys,ser,gly,phe,tyr,leu,lys,ile)
Five of the amino acids are converted to ketoglutarate
Some of these same amino acids can be converted
to citric acid cycle intermediates
Asparagine and aspartate are degraded to
oxaloacetate

Fate of Amino Acid Remnants

Those in pink can
represent precursors
for gluconeogenesis
Those in blue
do not yield glucose
if they produce
acetyl-CoA or
acetoacetyl-CoA