Crystallization and preliminary X-ray studies of the Ia component of Clostridium perfringens iota toxin complexed with NADPH

J Struct Biol. 1999 Jun 15;126(2):175-7. doi: 10.1006/jsbi.1999.4124.

Abstract

A recombinant Ia component of Clostridium perfringens iota toxin, which ADP-ribosylates actin, was crystallized by the hanging drop vapor diffusion method using PEG4000 as a precipitating agent. The crystals were obtained in the presence of NADPH, which is similar to a real substrate, NADH, and belongs to the space group P1 (a = 47.9 A, b = 54.5 A, c = 103.1 A, alpha = 99.0 degrees, beta = 93.3 degrees, and gamma = 107.2 degrees ). The Matthews coefficient of native crystal was 2.7, assuming 2 mol/asymmetric unit. Native data were collected at 2.4-A resolution. The results from a heavy-atom search showed that lanthanide ions (samarium, holmium) altered the molecular packing, judging from the unit-cell difference. The crystals also belonged to the space group P1 (a = 47.7 A, b = 53.9 A, c = 54.6 A, alpha = 68.9 degrees, beta = 78.3 degrees, and gamma = 73.7 degrees ), which is consistent with only one molecule per asymmetric unit.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADP Ribose Transferases*
  • Bacterial Proteins / chemistry
  • Bacterial Toxins / chemistry*
  • Clostridium perfringens / chemistry*
  • Crystallization
  • Crystallography, X-Ray
  • Microscopy, Polarization
  • NADP / chemistry*
  • Polyethylene Glycols / chemistry

Substances

  • Bacterial Proteins
  • Bacterial Toxins
  • iota toxin, Clostridium perfringens
  • Polyethylene Glycols
  • NADP
  • ADP Ribose Transferases