Cagayan State University - Andrews Campus, Tuguegarao City

CLINICAL CHEMISTRT 2 Laboratory | TTh, 5:00 PM - 8:00 PM

Instructor Status Notes

Wilson Leal Laranang, RMT, MSc Not started

GROUP 6 Date Submitted: 10/15/2022

GALANG, OLAND DEEMS B.
VILLANUEVA, KREZA LYN E.
VILLENA, JEIZEL CLAIRE G.

BSMLS-4 Rating: ________________

Activity No. 5
GAMMA-GLUTAMYL TRANSFERASE (GGT) DETERMINATION

1. Discuss the complete reaction and principle of the reference method for gamma-glutamyl
transferase (GGT) determination.
REACTION: The transfer of the -glutamyl group from peptides and
other molecules to an acceptor is catalyzed by -Glutamyl
Transferase. The substrate itself, a particular amino acid or
peptide, or even water can serve as the -glutamyl acceptor, in
which case straightforward hydrolysis occurs. Only peptides or
peptide-like compounds with a terminal glutamate residue
connected to the remaining portion of the substance by the
terminal (-γ-) carboxyl are capable of being broken down by the
enzyme. Compared to glycine or the tripeptide (gly-gly-gly),
glycylglycine is five times more efficient as an acceptor, although
nothing is known about the ideal characteristics of the acceptor
cosubstrate. Comparatively speaking, the simple hydrolysis
reaction is much slower than the peptidase transfer reaction. Figure 1. An example of a
(Rifai, et al., 2018, p. 419) reaction catalyzed by the
enzyme. From Tietz Textbook
of Clinical Chemistry and
PRINCIPLE: The reaction uses glycylglycine as the acceptor and
molecular Diagnostics (6th ed.,
L-glutamyl-3-carboxy-4-nitroanilide as the substrate. p. 420)
Glycylglycine itself acts as a buffer. The 5-amino-2-nitrobenzoate
reaction product's measured wavelength is 410 nm, and the reaction's temperature is 37 °C.
(Rifai, et al., 2018, p. 420-421)

Rifai, N. et al. (2018). Tietz Textbook of Clinical Chemistry and Molecular Diagnostics (6th ed.,

pp. 419-421). Elsevier Inc.

Cagayan State University - Andrews Campus, Tuguegarao City
CLINICAL CHEMISTRT 2 Laboratory | TTh, 5:00 PM - 8:00 PM
Instructor Status Notes

Wilson Leal Laranang, RMT, MSc Not started

2. Describe GGT: Its biochemistry and tissue distribution.

GGT gamma-glutamyl transpeptidase (EC 2.3.2.2) is a peptidase enzyme

It catalyzes the transfer of the -glutamyl group from peptides and other compounds containing
it to the substrate itself, some amino acid or peptide, or even water, where in case simple
hydrolysis occurs.

It cleaves glutathione (GSH) to allow cysteine recapture for intracellular GSH

production. GGT is involved in amino acid transport through the cellular membrane as
well as leukotriene metabolism (See Figure 1.).

It is necessary for glutathione metabolism, amino acid uptake, and oxidant defense.

Although GGT may be found in the cell membranes of many organs, including the kidneys,
bile duct, pancreas, gallbladder, spleen, heart, brain, and seminal vesicles, the liver is the
predominant source (primarily biliary epithelium).

GGT is present largely in the membrane and as aggregates in microsomes (from SER).

The cytoplasm contains a minor percentage (5%). Enhanced activity in serum is caused by
disaggregation (solubilization) and increased synthesis.

GGT is a particular indicator of cholestasis and biliary consequences in hepatobiliary

damage. It was discovered to be a particular marker of bile duct lesions in the rat liver.

Dzoyem, J. P. et al. (2014). Biochemical Parameters in Toxicological Studies in Africa.

Toxicological Survey of African Medicinal Plants, 659–715. https://doi.org/10.1016/b978-

0-12-800018-2.00023-6
Cagayan State University - Andrews Campus, Tuguegarao City
CLINICAL CHEMISTRT 2 Laboratory | TTh, 5:00 PM - 8:00 PM
Instructor Status Notes

Wilson Leal Laranang, RMT, MSc Not started

3. What is/are the clinical significance of GGT?

GGT is an enzyme that is present throughout the body, it is mostly located in the liver.
GGT may seep into the bloodstream if the liver is injured, thus, high levels of GGT in
the blood could be an indication of bile duct injury or liver disease. (U.S. National
Library of Medicine, n.d. para. 1)

GGT levels that are higher than usual could indicate liver damage. The damage may be
due to one of the following conditions: hepatitis, cirrhosis, alcohol use disorder,
pancreatitis, diabetes, congestive heart failure, side effect of a drug. Certain medicines
can cause liver damage in some people. (U.S. National Library of Medicine, n.d. para. 8)

The results are unable to identify the condition that is harming your liver. Typically, the
more GGT present, the more liver damage there will be. If your results show you have
low or normal levels of GGT, it means you probably don't have liver disease. (U.S.
National Library of Medicine, n.d. para. 9)

U.S. National Library of Medicine. (n.d.). Gamma-glutamyl transferase (GGT) test: Medlineplus

medical test. MedlinePlus.

https://medlineplus.gov/lab-tests/gamma-glutamyl-transferase-ggt-test/#:~:text=A

%20gamma%2Dglutamyl%20transferase%20(GGT)%20test%20measures%20the

%20amount,damage%20to%20the%20bile%20ducts.

4. Describe other methods of GGT determination.

The substrate for early GGT experiments was L—glutamyl-p-nitroanilide (GGPNA), with
glycylglycine acting as the -glutamyl residue acceptor. However, considering GGPNA has a
low solubility in the reaction mixture, obtaining saturating substrate concentrations is
complicated. The yellow hue of the p-nitroaniline formed in the reaction is measured at 405
nm (Rifai et al., 2018, p. 420).
GGPNA derivatives are available and have been employed in different approaches. To
boost water solubility, different groups have been added into the benzene ring using these
derivatives. The most useful of these substrates is L—glutamyl3-carboxy-4-nitroanilide, which is
soluble in water and is split by GGT at a rate equivalent to GGPNA. The IFCC reference
measurement protocol for GGT uses L—glutamyl-3-carboxy-4-nitroanilide as the substrate and
glycylglycine as the acceptor. Glycylglycine alone provides buffering. The reaction temperature
is 37°C, and the wavelength of the reaction product, 5-amino-2-nitrobenzoate, is 410 nm.
Cagayan State University - Andrews Campus, Tuguegarao City
CLINICAL CHEMISTRT 2 Laboratory | TTh, 5:00 PM - 8:00 PM
Instructor Status Notes

Wilson Leal Laranang, RMT, MSc Not started

Because the carboxy derivative has a greater absorbance than the noncarboxylated substrate,
the longer wavelength decreases the blank absorbance (Rifai et al., 2018, pp. 420-421).

Rifai, N. et al. (2018). Tietz Textbook of Clinical Chemistry and Molecular Diagnostics (6th ed.,

pp. 420-421). Elsevier Inc.

5. Discuss the correlation of ALP and GGT tests.

Skeletal disease is not accompanied by elevated serum activity, so measuring GGT is
clinically useful in determining the cause of obscure ALP increases. (Rifai, et al., 2018, p. 420)
During pregnancy, serum levels of GGT are different from those of ALP because GGT stays
normal even when there is cholestasis (McPherson, et al., 2017, pp. 296-297). Liver disorders
cause an increase in both GGT and ALP, diseases that impact bone tissue will only cause an
increase in ALP. To ascertain if the higher ALP result is the consequence of a liver or bone
illness, GGT might be utilized as a follow-up to an elevated ALP (Gamma-Glutamyl Transferase
(GGT), 2021).
While an ALP test may be done alone as part of a standard liver panel to screen for liver
damage, a GGT test may also be requested when findings of the ALP test are high but those of
the other tests that make up the liver panel (such as AST and ALT) are not elevated. Results
are evaluated in conjunction with each enzyme level and any other tests that may have been
run simultaneously. An elevated ALP with a high GGT level helps rule out bone disease as the
cause of the increased ALP level; elevated ALP with a low or normal GGT means the increased
ALP is more likely due to bone disease (Gamma-Glutamyl Transferase (GGT), 2021).

Gamma-Glutamyl Transferase (GGT). (November, 2021). TESTING.COM.

https://www.testing.com/tests/gamma-glutamyl-transferase-ggt/#:~:text=Both%20ALP

%20and%20GGT%20are,is%20most%20likely%20bone%20disease.

Rifai, N. et al. (2018). Tietz Textbook of Clinical Chemistry and Molecular Diagnostics (6th ed.,

p.

420). Elsevier Inc.

McPherson, R. A. & Pincus M. R. (2017). Henry’s Clinical Diagnosis and Management by

Laboratory Methods. (23rd ed., pp. 296-297). Elsevier Inc.

Cagayan State University - Andrews Campus, Tuguegarao City
CLINICAL CHEMISTRT 2 Laboratory | TTh, 5:00 PM - 8:00 PM
Instructor Status Notes

Wilson Leal Laranang, RMT, MSc Not started

BIBLIOGRAPHY

Dzoyem, J. P. et al. (2014). Biochemical Parameters in Toxicological Studies in Africa.

Toxicological Survey of African Medicinal Plants, 659–715. https://doi.org/10.1016/b978-

0-12-800018-2.00023-6

Gamma-Glutamyl Transferase (GGT). (November, 2021). TESTING.COM.

https://www.testing.com/tests/gamma-glutamyl-transferase-ggt/#:~:text=Both%20ALP

%20and%20GGT%20are,is%20most%20likely%20bone%20disease.

McPherson, R. A. & Pincus M. R. (2017). Henry’s Clinical Diagnosis and Management by

Laboratory Methods. (23rd ed., pp. 296-297). Elsevier Inc.

Rifai, N. et al. (2018). Tietz Textbook of Clinical Chemistry and Molecular Diagnostics (6th ed.,

pp. 419-421). Elsevier Inc.

U.S. National Library of Medicine. (n.d.). Gamma-glutamyl transferase (GGT) test: Medlineplus

medical test. MedlinePlus.

https://medlineplus.gov/lab-tests/gamma-glutamyl-transferase-ggt-test/#:~:text=A

%20gamma%2Dglutamyl%20transferase%20(GGT)%20test%20measures%20the

%20amount,damage%20to%20the%20bile%20ducts.