Frq2 from Drosophila melanogaster: cloning, expression, purification, crystallization and preliminary X-ray analysis.
Drosophila melanogaster contains two calcium-binding proteins, Frq1 and Frq2, in the nervous system that control the number of synapses and the probability of release. To understand the differential function of the two proteins, whose sequence is only 5% dissimilar, the crystal structures of Frq1 and Frq2 are needed. Here, the cloning, expression, purification, crystallization and preliminary crystallographic analysis of Frq2 are presented. The full-length protein was purified using a two-step chromatographic procedure. Two different diffracting crystal forms were obtained using a progressive streak-seeding method and detergents.
Department of Crystallography and Structural Biology, Institute of Physical Chemistry `Rocasolano', CSIC, Serrano 119, 28006 Madrid, Spain.
Soledad
S
Baños-Mateos
Department of Crystallography and Structural Biology, Institute of Physical Chemistry `Rocasolano', CSIC, Serrano 119, 28006 Madrid, Spain.
Antonio
A
Chaves-Sanjuán
Department of Molecular, Cellular and Developmental Neurobiology, Institute Cajal, CSIC, Avenida Dr Arce 37, 28002 Madrid, Spain.
Alicia
A
Mansilla
Department of Molecular, Cellular and Developmental Neurobiology, Institute Cajal, CSIC, Avenida Dr Arce 37, 28002 Madrid, Spain.
Alberto
A
Ferrús
Department of Crystallography and Structural Biology, Institute of Physical Chemistry `Rocasolano', CSIC, Serrano 119, 28006 Madrid, Spain.
María José
MJ
Sánchez-Barrena
D000818:Animals; D002135:Calcium-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D003001:Cloning, Molecular; D003460:Crystallization / Q000379:methods*; D018360:Crystallography, X-Ray / Q000379:methods*; D029721:Drosophila Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D004331:Drosophila melanogaster / Q000378:metabolism*
D002135:Calcium-Binding Proteins; D029721:Drosophila Proteins; C527166:FRQ2 protein, Drosophila
Drosophila melanogaster; Frq2; calcium-binding proteins