Immobilized analogues of sunflower trypsin inhibitor-1 constitute a versatile group of affinity sorbents for selective isolation of serine proteases.
Sunflower trypsin inhibitor-1 (SFTI-1), a natural 14-residue cyclic peptide, and some of its synthetic acyclic variants are potent protease inhibitors displaying peculiar inhibitory profiles. Here we describe the synthesis and use of affinity sorbents prepared by coupling SFTI-1 analogues to agarose resin. Chymotrypsin- and trypsin-like proteases could then be selectively isolated from pancreatin; similarly, other proteases were obtained from distinct biological sources. The binding capacity of [Lys5]-SFTI-1-agarose for trypsin was estimated at over 10 mg/mL of packed gel. SFTI-1-based resins could find application either to improve the performance of current purification protocols or as novel protease-discovery tools in different areas of biological investigation.
Pereira
Hugo Juarez Vieira
HJ
Department of Biochemistry and Immunology, University of São Paulo, Faculty of Medicine of Ribeirão Preto, 14049-900 Ribeirão Preto, SP, Brazil.
Salgado
Maria Cristina Oliveira
MC
Oliveira
Eduardo Brandt
EB
D000818:Animals; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D006368:Helianthus / Q000737:chemistry*; D010194:Pancreatin / Q000737:chemistry; D010456:Peptides, Cyclic / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry*; D011485:Protein Binding; D012117:Resins, Synthetic / Q000737:chemistry; D012697:Serine Endopeptidases / Q000737:chemistry / Q000302:isolation & purification*; D013552:Swine; D014361:Trypsin Inhibitors / Q000737:chemistry*
D010456:Peptides, Cyclic; D010940:Plant Proteins; D012117:Resins, Synthetic; C448501:SFTI-1 peptide, sunflower; D014361:Trypsin Inhibitors; D010194:Pancreatin; D012697:Serine Endopeptidases