Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Comparative Study
. 2008 Apr 1;64(Pt 4):281-3.
doi: 10.1107/S1744309108005678. Epub 2008 Mar 21.

Cloning, expression, crystallization and preliminary X-ray data analysis of norcoclaurine synthase from Thalictrum flavum

Alessandra Pasquo  1 , Alessandra Bonamore, Stefano Franceschini, Alberto Macone, Alberto Boffi, Andrea Ilari
Affiliations
Comparative Study

Cloning, expression, crystallization and preliminary X-ray data analysis of norcoclaurine synthase from Thalictrum flavum

Alessandra Pasquo et al. Acta Crystallogr Sect F Struct Biol Cryst Commun. .

Abstract

Norcoclaurine synthase (NCS) catalyzes the condensation of 3,4-dihydroxyphenylethylamine (dopamine) and 4-hydroxyphenylacetaldehyde (4-HPAA) as the first committed step in the biosynthesis of benzylisoquinoline alkaloids in plants. The protein was cloned, expressed and purified. Crystals were obtained at 294 K by the hanging-drop vapour-diffusion method using ammonium sulfate and sodium chloride as precipitant agents and diffract to better than 3.0 A resolution using a synchrotron-radiation source. The crystals belong to the trigonal space group P3(1)21, with unit-cell parameters a = b = 86.31, c = 118.36 A. A selenomethionine derivative was overexpressed, purified and crystallized in the same space group. A complete MAD data set was collected at 2.7 A resolution. The model is under construction.

PubMed Disclaimer

Figures

Figure 1
Figure 1
SDS–PAGE of fractions eluted from the HisTrap Fast Flow column. Lane M, molecular-weight markers (kDa); lanes 1–9, HisTrap elution fractions at 0.144, 0.153, 0.250, 0.259, 0.269, 0.279, 0.288, 0.298 and 0.307 M imidazole, respectively.
Figure 2
Figure 2
A typical SeMet NCS crystal (crystal dimensions 0.3 × 0.3 × 0.2 mm).
Figure 3
Figure 3
X-ray diffraction of an SeMet NCS crystal to 2.7 Å at the peak wavelength (λ = 0.97966 Å). Data were collected as 1.0° oscillation frames at 100 K.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Bohle, B. (2007). Allergy, 62, 3–10. - PubMed
    1. Collaborative Computational Project, Number 4 (1994). Acta Cryst. D50, 760–763. - PubMed
    1. Cowtan, K. (1994). Jnt CCP4/ESF–EACBM Newsl. Protein Crystallogr.31, 34–38.
    1. Cowtan, K. D. & Main, P. (1996). Acta Cryst. D52, 43–48. - PubMed
    1. Ebner, C., Hoffmann-Sommergruber, K. & Breiteneder, H. (2001). Allergy, 56, 43–44. - PubMed

Publication types

MeSH terms