Bacterial expression, purification and preliminary X-ray crystallographic characterization of the invertase inhibitor Nt-CIF from tobacco

• Published in: Acta crystallographica. Section D, Biological crystallography. Vol. 59; no. 12; pp. 2279 - 2282
• Main Authors: Hothorn, Michael, Bonneau, Fabien, Stier, Gunter, Greiner, Steffen, Scheffzek, Klaus
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England Munksgaard International Publishers 01.12.2003
• Subjects: beta-Fructofuranosidase - antagonists & inhibitors; carbohydrate metabolism; Cloning, Molecular; Crystallography, X-Ray; Enzyme Inhibitors - chemistry; Enzyme Inhibitors - metabolism; Enzyme Inhibitors - pharmacology; Gene Expression; invertases; Nicotiana - chemistry; Nicotiana tabacum; pectin methylesterase; plant enzymes; Plant Proteins - biosynthesis; Plant Proteins - chemistry; Plant Proteins - genetics; Plant Proteins - pharmacology; post-translational regulation; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - pharmacology
• ISSN: 1399-0047; 0907-4449; 1399-0047
• DOI: 10.1107/S0907444903021036

Acid invertases catalyzing the breakdown of sucrose are regulated at the post‐translational level by extracellular inhibitory proteins of 16–­20 kDa molecular weight in a pH‐dependent manner. Little is known about the characteristics of the underlying protein–protein interaction. Here, the expression, purification, characterization, crystallization and initial X‐ray analysis of a biologically active invertase inhibitor Nt‐CIF from tobacco is reported. Four crystal forms covering a wide pH range have been obtained and data sets at resolutions higher than 2.5 Å have been collected.