1. Crystallization and preliminary X-ray diffraction studies of aSFP, a bovine seminal plasma protein with a single CUB domain architecture
- Author
-
Antonio A. Romero, Paloma F. Varela, João Miguel L. Dias, Maria João Romão, Ana Luísa Carvalho, Ingo Kölln, Edda Töpfer-Petersen, Juan J. Calvete, and Claus Urbanke
- Subjects
Molecular mass ,Seminal Plasma Proteins ,Resolution (electron density) ,CUB domain ,Biochemistry ,law.invention ,chemistry.chemical_compound ,Crystallography ,Monomer ,chemistry ,law ,X-ray crystallography ,Molecule ,Crystallization ,Molecular Biology - Abstract
Bovine acidic seminal fluid protein (aSFP) is a 1.29 kDa polypeptide of the spermadhesin family built by a single CUB domain architecture. The CUB domain is an extracellular module present in 16 functionally diverse proteins. To determine the three-dimensional structure of aSFP, the protein was crystallized at 21 degrees C by vapor diffusion in hanging drops, using ammonium sulfate, pH 4.7, and polyethyleneglycol 4,000 as precipitants, containing 10% dioxane to avoid the formation of clustered crystals. Elongated prismatic crystals with maximal size of 0.6 x 0.3 x 0.2 mm3 diffract to beyond 1.9 A resolution and belong to space group P2(1)2(1)2(1), with cell parameters a = 52.4 A, b = 41.5 A, c = 48.2 A. There is one aSFP molecule per asymmetric unit, which corresponds to a crystal volume per unit molecular mass of 2.04 A3/Da, and analytical ultracentrifugation analysis show that aSFP is a monomeric protein.
- Published
- 1997
- Full Text
- View/download PDF