Crystallization and preliminary X-ray crystallographic studies on a Kunitz-type potato serine protease inhibitor.

E.A.J. Thomassen, L.A.M. Pouvreau, H. Gruppen, J.P. Abrahams

Research output: Contribution to journalArticleAcademicpeer-review

2 Citations (Scopus)

Abstract

Interest in protease inhibitors has been renewed because of their potent activity in preventing carcinogenesis in a wide variety of in vivo and in vitro model systems. Potato tubers contain a wide range of such protease inhibitors. In cv. Elkana potato tubers, protease inhibitors represent about 50% of the total amount of soluble protein. Potato serine protease inhibitor (PSPI), one of the isoforms of the most abundant group of protease inhibitors, is a dimeric double-headed Kunitz-type inhibitor. No high-resolution structural information on this type of inhibitor has so far been obtained, as all currently known structures are of the monomeric single-headed or monomeric double-headed types. Crystals were grown in 0.1 M HEPES pH 7.5, 10% PEG 8000 and 8% ethylene glycol complemented with 9 mM 1-s-octyl-beta-D-thioglucoside or 0.1 M glycine. Data were collected from a single crystal under cryoconditions to 1.8 Å resolution. The protein crystallized in space group P21, with unit-cell parameters a = 54.82, b = 93.92, c = 55.44 Angstrom, beta = 100.7 °; the scaling Rsym is 0.044 for 45 456 unique reflections.
Original languageEnglish
Pages (from-to)1464-1466
JournalActa Crystallographica Section D-Biological Crystallography
Volume60
Issue number8
DOIs
Publication statusPublished - 2004

Keywords

  • molecular replacement
  • cv elkana

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