Enzyme localization
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Biological catalysts, called enzymes are invariably proteins, which catalyze specific biochemical reactions in cells and biological tissues of body. Different tissues and cells may be rich in certain specific enzymes. Inside cells, different cell organelles like cell nuclei, nucleoli, mitochondria, endoplasmic reticulum, Golgi body, lysosomes, chloroplasts, etc contain different sets and subsets of enzymes. The enzyme localization can be studied,
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either in (1) separated purified fractions of the cell by differential centrifugation, or ultracentrifugation techniques or (2) in tissue sections by micro-spectrophotometry. Specific enzyme substrate (biology) is used to study chosen enzyme(s). Purified cell fraction containing one type of cell organelles are incubated with a requisite amount of substrate at a fixed temperature for a fixed time period. The product of the enzyme action is measured with a spectrophotometer or with a colorimeter (chemistry) after doing a procedurally specified set of chemical reactions to make the product colored.[1] For studying enzyme activity in tissue sections, unfixed frozen sections of tissue, cut with a freezing microtome/cryostat are bathed in substrate solution at a requisite temperature for a requisite time. The product of enzyme action is converted into a colored or black/grey product and its histology is viewed under microscope with or without fixation (histology) for localization of enzyme activity (Fig. 1).[2]
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References[edit]
- ↑ Bradfield J R G (January 1950) The localization of enzymes in cells. Biological Reviews, vol. 25 (No. 1), pp. 113-157.http://onlinelibrary.wiley.com/doi/10.1111/j.1469-185X.1950.tb00588.x/abstract
- ↑ Bawa S R and YashRoy R C (1972) Vulture retina enzyme distribution and function. Neurobioloy, vol.2 No.4) pp.162-168. https://www.academia.edu/7707436/Bawa_S_R_and_YashRoy_R_C_1972_Vulture_retina_enzyme_distribution_and_function._Neurobiology._Vol._2_No._4_pp._162-168
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