Expression, purification, crystallization and preliminary X-ray analysis of the cathelicidin motif of the protegrin-3 precursor.
Numerous precursors of antibacterial peptides with unrelated sequences share a similar prosequence which belongs to the cathelicidin family of proteins. The three-dimensional structure of this cathelicidin motif, which contains two disulfide bonds, has not yet been reported. The cathelicidin motif (ProS) of the protegrin-3 precursor was overexpressed in Escherichia coli as a His-tagged protein. The His(6) tag was removed by thrombin cleavage. ProS was purified to homogeneity and single crystals were obtained by the hanging-drop vapour-diffusion method at pH 3-4. Preliminary X-ray diffraction analysis indicated that these crystals belong to the hexagonal space group P6(1)22 or P6(5)22, with unit-cell parameters a = b = 51.42, c = 134.25 A. These crystals diffracted beyond 2.75 A (1.9 A at ESRF) and contain one molecule per asymmetric unit.
Sanchez
J F
JF
Centre de Biochimie Structurale, UMR 5048 CNRS-UM1/UMR 554 INSERM-UM1, Université Montpellier 1, Faculté de Pharmacie, 15 Avenue Charles Flahault, 34060 Montpellier CEDEX 2, France.
Hoh
F
F
Strub
M P
MP
Strub
J M
JM
Van Dorsselaer
A
A
Lehrer
R
R
Ganz
T
T
Chavanieu
A
A
Calas
B
B
Dumas
C
C
Aumelas
A
A
D020816:Amino Acid Motifs; D023181:Antimicrobial Cationic Peptides; D001798:Blood Proteins / Q000737:chemistry; D003460:Crystallization; D018360:Crystallography, X-Ray; D011487:Protein Conformation; D011498:Protein Precursors / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry
D023181:Antimicrobial Cationic Peptides; D001798:Blood Proteins; D011498:Protein Precursors; D011506:Proteins; D011994:Recombinant Proteins; C121626:cathelicidin 3 protein, Equus caballus; C094881:protegrin-3