A Method for Urinary Trypsin Inhibitor (UTI) Purification Combining Anion-Exchange Chromatography Enrichment and Preparative SDS-PAGE

Gabriele Nieddu; Marilena Formato; Antonio Junior Lepedda

doi:10.1007/978-1-0716-2946-8_17

A Method for Urinary Trypsin Inhibitor (UTI) Purification Combining Anion-Exchange Chromatography Enrichment and Preparative SDS-PAGE

Methods Mol Biol. 2023:2619:239-248. doi: 10.1007/978-1-0716-2946-8_17.

Authors

Gabriele Nieddu¹, Marilena Formato¹, Antonio Junior Lepedda²

Affiliations

¹ Department of Biomedical Sciences, University of Sassari, Sassari, Italy.
² Department of Biomedical Sciences, University of Sassari, Sassari, Italy. ajlepedda@uniss.it.

PMID: 36662474
DOI: 10.1007/978-1-0716-2946-8_17

Abstract

This chapter describes a method for the purification of urinary trypsin inhibitor (UTI), a small chondroitin sulfate proteoglycan with Ser-proteinase inhibitory activity, excreted at high levels into urine following an inflammatory condition. The method consists of two fractionation steps: an anion-exchange chromatography and a sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by Coomassie Brilliant Blue G-250 gel staining. Several UTI bands are excised from gel, minced, destained, and dehydrated for extraction with SDS-containing buffer, at 60 °C for 24 h. This allows for obtaining a highly purified UTI sample useful for both structural and functional studies.

Keywords: Anion-exchange chromatography; Chondroitin sulfate proteoglycan; Preparative SDS-PAGE; Ulinastatin; Urinary trypsin inhibitor.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Anions
Chondroitin Sulfate Proteoglycans*
Chromatography*
Electrophoresis, Polyacrylamide Gel
Trypsin

Substances

urinastatin
Chondroitin Sulfate Proteoglycans
Anions
Trypsin