Protein Sequencing - Determining Amino Acid Composition

Determining Amino Acid Composition

It is often desirable to know the unordered amino acid composition of a protein prior to attempting to find the ordered sequence, as this knowledge can be used to facilitate the discovery of errors in the sequencing process or to distinguish between ambiguous results. Knowledge of the frequency of certain amino acids may also be used to choose which protease to use for digestion of the protein. A generalized method often referred to as amino acid analysis for determining amino acid frequency is as follows:

  1. Hydrolyse a known quantity of protein into its constituent amino acids.
  2. Separate the amino acids in some way.

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