Details

Autor(en) / Beteiligte

• Li, Ying
• Yan, Youwei
• Zugay-Murphy, Joan
• Xu, Bei
• Cole, James L.
• Witmer, Marc
• Felock, Peter
• Wolfe, Abigail
• Hazuda, Daria
• Sardana, Mohinder K.
• Chen, Zhongguo
• Kuo, Lawrence C.
• Sardana, Vinod V.

Titel

Purification, solution properties and crystallization of SIV integrase containing a continuous core and C-terminal domain

Ist Teil von

• Acta crystallographica. Section D, Biological crystallography., 1999-11, Vol.55 (11), p.1906-1910

Ort / Verlag

5 Abbey Square, Chester, Cheshire CH1 2HU, England: International Union of Crystallography

Erscheinungsjahr

1999

Quelle

Wiley Online Library - AutoHoldings Journals

Beschreibungen/Notizen

• The C‐terminal two‐thirds segment of integrase derived from the simian immunodeficiency virus has been cloned, expressed in Escherichia coli, and purified to greater than 95% homogeneity. The protein encompasses amino‐acid residues 50–293 and contains a F185H substitution to enhance solubility. In dilute solutions at concentrations below 1 mg ml−1, the enzyme is predominantly dimeric. At the higher concentrations (>10 mg ml−1) required to enable crystallization, the enzyme self‐associates to form species with molecular weights greater than 200 kDa. Despite the apparent high aggregation in solution, the enzyme crystallizes from a 8%(v/v) polyethylene glycol (molecular weight 6000) solution in a form suitable for X‐ray diffraction studies. The resulting single crystals belong to the space group P212121, with unit‐cell parameters a = 79.76, b = 99.98, c = 150.2 Å, α = β = γ = 90° and Z = 4. Under X‐ray irradiation generated with a rotating‐anode generator, the crystals diffract to 2.8 Å resolution and allow collection of a native 3 Å resolution diffraction data set.