The application of 0.1 M quadrol to the microsequence of proteins and the sequence of tryptic peptides.
In an effort to extend automated Edman degradation to nanomole quantities of protein, the method of sequenator analysis described by Edman and Begg (Edman, P., and Begg, G. (1967), Eur. J. Biochem. 1, 80) was modified to permit long degradations in the absence of carrier proteins. By using an aqueous 0.1 M Quadrol program with limited, combined benezene-ethyl acetate solvent extractions, as well as a change in the delivery system for heptafluorobutyric acid, it was possible to recover and identify the first 30 amino acid residues from a sequenator run on 7 nmol of myoglobin. For 3 nmol of myoglobin, 20 steps could be identified. PTH-amino acids were identified by gas-liquid chromatography and thin-layer chromatography on polyamide sheets. Without using a carrier protein the cup to prevent mechanical losses (Niall, H. D., Jacobs, J. W., Van Rietshoten, J., and Tregear, G. W. (1974), FEBS Lett. 41, 62), the repetitive yield using this program was 93-96%. The same program has been applied successfully to peptides of 14 or more residues with or without modification by Braunitzer's reagent and to a number of larger peptides and proteins including a 216 residue segment of rabbit antibody heavy chain in which a sequence of 35 steps was accomplished on 25 nmol.
D000595:Amino Acid Sequence; D001322:Autoanalysis / Q000379:methods*; D002845:Chromatography; D002855:Chromatography, Thin Layer; D005029:Ethylenediamines; D007128:Immunoglobulin Fragments / Q000032:analysis; D008722:Methods; D009211:Myoglobin / Q000032:analysis; D010446:Peptide Fragments / Q000032:analysis*; D010669:Phenylthiohydantoin / Q000494:pharmacology; D011412:Propanolamines; D011506:Proteins / Q000032:analysis*
D005029:Ethylenediamines; D007128:Immunoglobulin Fragments; D009211:Myoglobin; D010446:Peptide Fragments; D011412:Propanolamines; D011506:Proteins; C010629:1,1',1'',1'''-(ethylenedinitrilo)tetra-2-propanol; D010669:Phenylthiohydantoin