2. Different techniques to establish structures
Several biochemical, chemical and biophysical techniques are available to determine the different structural levels of an enzyme after purification. If these are well established, robotic and computerized tools can save considerable time and increase precision. The most common techniques to establish these structures are shown below.
2.1 Primary sequences: total, limited proteolysis, Edman sequencing and RT-PCR
To establish the primary sequence of a purified enzyme, the sample must be treated so that all covalent bonds (essentially disulfide bridges) involved in the tertiary structure are broken.
Once the enzymatic protein has been “linearized”, its overall amino acid composition can be determined. The protein solution is usually...
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Different techniques to establish structures
Structure and function of three enzymes
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