PBPN (ANT2004)

TUTORIAL 8: PROTEIN METABOLISM

Name:_________________________________ Tutorial Class: ________ Date:__________

TASK 1: Transamination, Oxidative Deamination & Transdeamination

Describe transdeamination reactions. (AY1416, Apr, SA)
Use the following schematic representation to help you present your answers.
In transdeamination, the amino group from an amino acid is transferred to a alpha-
ketoacid/carbon-skeleton, the amino acid that loses the amino group becomes a alpha-
ketoacid/carbon-skeleton. While the alpha-ketoacid/carbon-skeleton receives the amino
group becomes an amino acid. In the representation, the amino acid alanine loses its amino
group to the alpha-ketogluterate to become pyruvate while alpha-ketogluterate becomes a
new amino acid, glutamate. This reversible reaction takes place in the cytosol and
mitochondria of all cells. The enzymes involved are transaminase(in the representation, the
enzyme alanine aminotransferase). Most reactions use alpha-ketogluterate as the amino
group acceptor (alpha-keto acid/carbon-skeleton) generating a common product glutamate.
In oxidative deamination, the amino group is removed from glutamate generating
ammonium ion and alpha-ketoglutarate. The reaction takes place in the mitochondrial matrix
of all cells. The enzyme involved is glutamate-dehydrogenase involving the coenzyme
NAD+.
Oxidative is always glutamate throw away amino group.
H3N+-CH-(CH3)-COO- O=C-(CH2-CH2-COO-)-
COO-
Enzyme: glutamate
E Nzyme: alanine aminotransferase dehydrogenase

ALANINE + α-KETOGUTARATE NH4+

PYRUVATE + GLUTAMATE O=C-(CH3)-COO-

H3N+-CH-(CH2-CH2-COO-)-COO-
TASK 2: Reviewing mechanisms of NH3 transport
Explain these TWO main mechanisms involved in the transport of NH3 to the liver for
excretion: (AY12/13Apr, main)
• Glucose alanine cycle
• Glutamine synthase/glutaminase system
Use the schematic representation in Appendix 1 to help you present your answers.
Identify the metabolic pathways a, b, c, d, e and f
Name the enzymes g, h, i and j
Can draw when come out in exam instead of writing
Common mistake in exam:
 When exam ask how muscle don’t draw other tissues only liver and muscle (alanine
moves so always draw alanine 1st)
A: transamination
B: transamination
C: oxidative deamination
D: gluconeogenesis
E: glycolysis
F:gluconeolysis
G: alanine aminotransferase
H: alanine aminotransferase
I: glutamine synthase
J: glutaminase

TASK 3: Clinical Application Question

(a). Complete the urea cycle given in Appendix 2.

Identify:
Cellular sites a and b
Enzymes c, d, e, f and g
Substrates/products h, i, j and k
No drawing only MCQ for urea cycle
A: mitochondrial matrix
B: cytosol
C: carbonyl phosphate synthetase
D: ornithine transcarbamoylase
E: arginosuccinate synthetase
F: argininosuccinase
G: arginase
H: carbamoyl phosphate
I: citrulline
J: argininosuccinate
K: ornithine
 • Besides the NH4+, ONE other source of NH3 is aspartate

• Fumarate that is generated is part of the TCA cycle and forms malate which gets
oxidized to form oxaloacetate

• Ornithine is regenerated and transported back to the mitochondrial matrix to start

another cycle of urea synthesis.
• Urea that is generated is brought to the kidney to be excreted in the urine

(b). Write an equation to represent the overall reaction in the urea cycle:

NH4+ + HCO3- + aspartate + 3ATp -> urea + fumarate + 2ADP + AMP + 4Pi

(c). The management of the rare urea cycle disorder, ornithine transcarbamoylase deficiency,
requires protein restriction as well as citrulline supplementation. With reference to (a),
explain how these two strategies work.
The entry point for NH3 in the urea cycle involves the formation of carbamoyl phosphate.
Ornithine transcarbamoylase then catalyses the conversion of carbamoyl phosphate to
citrulline. For patients with this enzyme deficiency, NH3’s entry into the urea cycle is
affected. Excess dietary protein is the source of NH3 as they are not stored but catabolized
this protein restriction is necessary to reduce NH3 load. Another entry point for NH3 into the
urea cycle is via aspartate. The provision of citrulline allows the entry of aspartate to form
urea, which helps in the excretion of NH3.