Methodology for purification of large hydrophobic peptides by high-performance liquid chromatography.
To aid in structural studies of pig cardiac myosin light chains (L27 and L28), a procedure of ion-exchange chromatography (IEC) on Trisacryl M (noted for its high capacity) in combination with reversed-phase high-performance liquid chromatography (RP-HPLC) and volatile buffers has been developed. In contrast with other IEC methods (resins or HPIEC), the use of Trisacryl M facilitates subsequent peptide purifications by RP-HPLC. The advantage of the present combination of techniques is also that it enables the isolation of hydrophobic peptides in high yield, e.g., the N-terminal chymotryptic peptide from L27 was thus purified and, after sub-digestion with trypsin, its sequence has been established.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D002918:Chymotrypsin / Q000494:pharmacology; D006868:Hydrolysis; D009206:Myocardium / Q000032:analysis*; D015879:Myosin Subfragments; D009218:Myosins / Q000302:isolation & purification*; D010446:Peptide Fragments / Q000302:isolation & purification*; D010455:Peptides / Q000302:isolation & purification; D013552:Swine
D000596:Amino Acids; D015879:Myosin Subfragments; D010446:Peptide Fragments; D010455:Peptides; D002918:Chymotrypsin; D009218:Myosins