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Advanced Protein Sequencing Techniques and Equipment

Learn about protein sequencing, endoproteinases, alignment, and spectroscopy methods using an Applied Biosystem Procise 492 peptide sequenator with Edman degradation. Discover the step-by-step process and timeline for sequencing a 50-amino acid residue peptide.

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Advanced Protein Sequencing Techniques and Equipment

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  1. Proteina intatta 1 2 3 4 Ala-Leu-Thr-Pro ... ... ... ... Endoproteinasi Arg-C 1 2 3 4 5 6 7 8 9 10 11 12 a) Ile-Lys-Val-Trp-Glu-Gly-Thr-Gly-Leu-Val-Pro-Arg 1 2 3 b) Ser-Thr-Val 1 2 3 4 5 6 7 8 9 10 c) Ala-Leu-Thr-Pro-Val-Ser-Glu-Met-Val-Arg Endoproteinasi Glu-C 1 2 3 4 5 6 7 8 9 10 d) Gly-Thr-Gly-Leu-Val-Pro-Arg-Ser-Thr-Val 1 2 3 4 5 6 7 e) Ala-Leu-Thr-Pro-Val-Ser-Glu 1 2 3 4 5 6 7 8 f) Met-Val-Arg-Ile-Lys-Val-Trp-Glu Allineamento per sovrapposizione zone comuni: e) Ala-Leu-Thr-Pro-Val-Ser-Glu c) Ala-Leu-Thr-Pro-Val-Ser-Glu-Met-Val-Arg f) Met-Val-Arg-Ile-Lys-Val-Trp-Glu a) Ile-Lys-Val-Trp-Glu-Gly-Thr-Gly-Leu-Val-Pro-Arg d) Gly-Thr-Gly-Leu-Val-Pro-Arg-Ser-Thr-Val b) Ser-Thr-Val Sequenza:  1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 Ala-Leu-Thr-Pro-Val-Ser-Glu-Met-Val-Arg-Ile-Lys-Val-Trp-Glu-Gly-Thr-Gly-Leu-Val-Pro-Arg-Ser-Thr-Val ____________________________ ___________ ____________________ ___________________________ ___________

  2. Anno Quantita' Tempo 196O 10 g 3 anni 1970 1 g 1 anno 1980 1 mg 1 settimana 1990 0.05 mg 1 giorno 2000 0.02 mg 1 giorno Quantita' e tempi richiesti negli anni per sequenziare un peptide di 50 residui amminoacidici

  3. Sequenziatore automatico di peptidi Applied Biosystem Procise 492 che opera secondo il metodo di Edman della degradazione con fenilisotiocianato

  4. Schema di un sequenziatore di peptidi

  5. Spettrometro di massa

  6. Standard PTH-a.acidi Degradazione di Edman

  7. Primo passo degradazione Edman Peptide singolo

  8. Secondo passo degradazione Edman Peptide singolo

  9. Degradazione di Edman su miscela di peptidi Tre passi

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