Collision Induced Dissociation to Detect the Amino Acid Sequence of Synthetic Peptide: A Tandem Mass Spectrometry Investigation
Disciplines
Analytical Chemistry
Abstract (300 words maximum)
Collision Induced fragmentation (CID) coupled with mass spectrometry can be implemented for structure elucidation of synthetic peptides. CID could be useful in determining the exact sequence for a peptide or a protein by observing the peaks formed due to b and y ions. The position of these peaks would correspond to mass/charge ratio. Comparing these peak values with masses of amino acids could deduce the sequence. In this experiment, synthetic Temporin L analogue was used as the sample peptide. Different collision energies were used ranging from 10-35 to observe the pattern of fragmentation over increasing normalized energy. As the normalized collision energy was increased, higher intensity peaks were observed showing more fragmentation of the peptide. Major peaks with higher abundance were observed and noted. At lower collision energy of 10, seven b and y ions are detected in which y ions are prevalent. No neutral loss was observed from the fragment ion when low collision energy was applied. When normalized collision energy was set to 30, twelve fragment ions identified. Among them, few b ions are produced, however, more y ions are generated. Most of the y ions produced with the neural loss of NH3. Such peaks are observed at m/z 1249.92 and 1378.00 can be assigned to y10-NH3 and y11-NH3, respectively may be due to the presence of two basic amino acids such as K and R.
Academic department under which the project should be listed
Chemistry and Biochemistry
Primary Investigator (PI) Name
Mohammad A. Halim
Collision Induced Dissociation to Detect the Amino Acid Sequence of Synthetic Peptide: A Tandem Mass Spectrometry Investigation
Collision Induced fragmentation (CID) coupled with mass spectrometry can be implemented for structure elucidation of synthetic peptides. CID could be useful in determining the exact sequence for a peptide or a protein by observing the peaks formed due to b and y ions. The position of these peaks would correspond to mass/charge ratio. Comparing these peak values with masses of amino acids could deduce the sequence. In this experiment, synthetic Temporin L analogue was used as the sample peptide. Different collision energies were used ranging from 10-35 to observe the pattern of fragmentation over increasing normalized energy. As the normalized collision energy was increased, higher intensity peaks were observed showing more fragmentation of the peptide. Major peaks with higher abundance were observed and noted. At lower collision energy of 10, seven b and y ions are detected in which y ions are prevalent. No neutral loss was observed from the fragment ion when low collision energy was applied. When normalized collision energy was set to 30, twelve fragment ions identified. Among them, few b ions are produced, however, more y ions are generated. Most of the y ions produced with the neural loss of NH3. Such peaks are observed at m/z 1249.92 and 1378.00 can be assigned to y10-NH3 and y11-NH3, respectively may be due to the presence of two basic amino acids such as K and R.