Glucose-6-Phosphate Dehydrogenase Activity
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|Office:||6th floor, Radiopoetro|
|Pre-test:||3-6 questions, short-answer.|
|Post-test:||Sometimes. Oral only.|
To find out the activity of the enzyme G-6-PD by measuring the increase in NADPH, which is measured by the increased absorbency of NADPH under UV light with a wavelength of 340 nm.
Functions of the Reagents
- Hemolysate: lysed erythrocyte that helps obtain enzyme G-6-PD.
- Triethanolamine: acts as buffer (maintains pH at: 7.35-7.45).
- EDTA: anti-coagulant, metal ions binder.
- NADP: NADP donor. Accepts hydrogen from glucose-6-phosphate.
- Glucose-6-phosphate (substrate): Converted to 6-phosphogluconate by enzyme G-6-PD.
- Incubation at 37°C: Optimal enzyme activity happens at 37°C (body temperature).
- Vortex: Hemogenization.
- Spectrophotometry: Provides UV rays at wavelength 340nm, the optimal wavelength for NADPH activity.
Batch 2011 Pre-test
- The function of NADPH is? Reduces oxidized-glutathione to reduced-glutathione, which protects erythrocytes against oxidative attacks.
- Glucose-6-phosphate dehydrogenase reaction? Glucose-6-phosphate + NADP → 6-Phosphogluconate + NADPH.
- The diseases caused by G-6-PD deficiency are? Neonatal hyperbilirubinemia, acute hemolysis, and chronic hemolysis.
- The decrease in reduced-glutathione leads to? Hemolytic anemia.
- The oxidant drugs that cause decreased reduced-glutathione concentration are? Quinine, analgesics, and sulfonamides.