Synthesis, purification, and chemical characterization of the amino-terminal 1-34 fragment of bovine parathyroid hormone synthesized by the solid-phase procedure.
Peptides prepared by solid-phase synthesis during a systematic study of structure-activity relations in parathyroid hormone have been subjected, after sequential purifications, to rigorous analysis of purity by a variety of analytical techniques including sequence analysis by the Edman procedure. The present paper undertakes a critical appraisal of the utility of different tests of peptide purity and the use of the procedures in guiding and monitoring optimal synthesis strategies. Sequence analysis of a peptide representing the amino-terminal 34 residues of bovine parathyroid hormone (bPTH-(1-34)) revealed the presence of at least 30% of contaminating error peptides which were undetected by other analytical procedures. The major contaminant was identified as a peptid in which glutamine at position 29 was deleted. A repeat synthesis using fluorescamine rather than ninhydrin to monitor the coupling reaction resulted in a preparation that lacked the contaminant resulting from deletion. These findings demonstrate the particular value of sequence analysis in the evaluation of purity of peptides synthesized by the solid-phase technique.
D000262:Adenylyl Cyclases / Q000378:metabolism; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001681:Biological Assay; D002118:Calcium / Q000097:blood; D002417:Cattle; D055598:Chemical Phenomena; D002621:Chemistry; D002645:Chickens; D004789:Enzyme Activation; D010281:Parathyroid Hormone / Q000138:chemical synthesis*; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000302:isolation & purification / Q000494:pharmacology; D013329:Structure-Activity Relationship
D000596:Amino Acids; D010281:Parathyroid Hormone; D010446:Peptide Fragments; D000262:Adenylyl Cyclases; D002118:Calcium