Crystallization of the C-terminal globular domain of avian reovirus fibre.
Avian reovirus fibre, a homotrimer of the sigmaC protein, is responsible for primary host-cell attachment. Using the protease trypsin, a C-terminal sigmaC fragment containing amino acids 156-326 has been generated which was subsequently purified and crystallized. Two different crystal forms were obtained, one grown in the absence of divalent cations and belonging to space group P6(3)22 (unit-cell parameters a = 75.6, c = 243.1 A) and one grown in the presence of either zinc or cadmium sulfate and belonging to space group P321 (unit-cell parameters a = 74.7, c = 74.5 A and a = 73.1, c = 69.9 A for the Zn(II)- and Cd(II)-grown crystals, respectively). The first crystal form diffracted synchrotron radiation to 3.0 A resolution and the second form to 2.2-2.3 A. Its closest related structure, the C-terminal fragment of mammalian reovirus fibre, has only 18% sequence identity and molecular-replacement attempts were unsuccessful. Therefore, a search is under way for suitable heavy-atom derivatives and attempts are being made to grow protein crystals containing selenomethionine instead of methionine.
van Raaij
Mark J
MJ
Departamento de Bioquímica y Biología Molecular, Facultad de Farmacia, Universidad de Santiago de Compostela, Campus Sur, E-15782 Santiago de Compostela, Spain. vanraaij@usc.es
Hermo Parrado
X Lois
XL
Guardado Calvo
Pablo
P
Fox
Gavin C
GC
Llamas-Saiz
Antonio L
AL
Costas
Celina
C
Martínez-Costas
José
J
Benavente
Javier
J
D019187:Cadmium Compounds / Q000737:chemistry; D002213:Capsid / Q000737:chemistry*; D002412:Cations; D003460:Crystallization; D018360:Crystallography, X-Ray; D004247:DNA / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D008715:Methionine / Q000737:chemistry; D030101:Orthoreovirus, Avian / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D012645:Selenomethionine / Q000737:chemistry; D013431:Sulfates / Q000737:chemistry; D019287:Zinc Sulfate / Q000737:chemistry
D019187:Cadmium Compounds; D002412:Cations; D013431:Sulfates; D019287:Zinc Sulfate; D004247:DNA; C037123:cadmium sulfate; D012645:Selenomethionine; D008715:Methionine