Anaerobic crystallisation of an isopenicillin N synthase.Fe(II).substrate complex demonstrated by X-ray studies.

Roach, P; Clifton, I; Hensgens, C; Shibata, N; Long, A; Strange, R; Hasnain, S; Schofield, C; Baldwin, J; Hajdu, J

Journal article

Anaerobic crystallisation of an isopenicillin N synthase.Fe(II).substrate complex demonstrated by X-ray studies.

Abstract:: Isopenicillin N synthase (IPNS) was cocrystallised with ferrous sulphate and its substrate, delta-(L-alpha-aminoadipoyl)-L-cysteinyl-D-valine (Aad-Cys-Val). Vital to the successful procedure was the maintenance of a rigorously anaerobic environment. Hanging-drop vapour-diffusion crystallisation experiments, using lithium sulphate as the precipitant produced three crystal forms. Form I crystals, with a plate habit, diffracted X-rays to at least 0.11-nm resolution at the European Synchrotron Radiation Facility and belong to the space group P2(1)2(1)2(1), with unit-cell dimensions a = 4.68, b = 7.15, c = 10.10 nm. Their asymmetric unit contains a single IPNS.Fe(II).Aad-Cys-Val complex with a solvent content of 38.5%. Form II crystals, with a hexagonal habit, diffract X-rays to at least 0.21 nm resolution at the European Synchrotron Radiation Facility and belong to the space group P3(1)21, with unit-cell dimensions a = 10.10, b = 10.10, c = 11.567 nm. Their asymmetric unit also contains a single IPNS.Fe(II).Aad-Cys-Val complex with a solvent content of 69.5%. Form III crystals, needles, do not show well-ordered diffraction. Although all three forms were initially produced in crystallisation experiments under identical conditions, appropriate micro and streak seeding allows selective crystallisation of form I or form II crystals. Extended X-ray-absorption fine-structure studies on a crystalline slurry of the form I crystals demonstrate the presence of an Fe-S(Aad-Cys-Val) bond length of 0.234 +/- 0.003 nm.

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APA Style

Roach, P., Clifton, I., Hensgens, C., Shibata, N., Long, A., Strange, R., Hasnain, S., Schofield, C., Baldwin, J., & Hajdu, J. (1996). Anaerobic crystallisation of an isopenicillin N synthase.Fe(II).substrate complex demonstrated by X-ray studies. European Journal of Biochemistry / FEBS, 242(3), 736–740.

MLA Style

Roach, P., et al. “Anaerobic Crystallisation of an Isopenicillin N Synthase.Fe(II).Substrate Complex Demonstrated by X-Ray Studies.” European Journal of Biochemistry / FEBS, vol. 242, no. 3, 1996, pp. 736–40.

Chicago Style

Roach, P, I Clifton, C Hensgens, N Shibata, A Long, R Strange, S Hasnain, C Schofield, J Baldwin, and J Hajdu. 1996. “Anaerobic Crystallisation of an Isopenicillin N Synthase.Fe(II).Substrate Complex Demonstrated by X-Ray Studies.” European Journal of Biochemistry / FEBS 242 (3): 736–40.
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