The binding of calcium to the B-repeat segment of SdrD, a cell surface protein of Staphylococcus aureus.
In the Sdr family of Staphylococcus aureus cell surface proteins, three recently cloned members (Josefsson, E., McCrea, K., Ni Eidhin, D., O'Connell, D., Cox, J. A., Hook, M., and Foster, T. (1998) Microbiology, in press) display variable numbers of B-repeats, i.e. segments of 110-113 residues that probably make up one folding unit. Each B-repeat contains one conserved EF-hand motif and two acidic stretches. Equilibrium dialysis revealed that segment B1-B5 of SrdD contains 14 Ca2+-binding sites with high affinity ([Ca2+]0.5, 4 microM), whereas flow dialysis yielded 5 sites of high affinity (class I) and 10 of low affinity (class II). The discrepancy could be explained by the slow induction of high affinity in the class II sites. Kinetic experiments using fluorescent Ca2+ indicators corroborated slow binding of Ca2+ at the latter sites. Circular dichroism and Trp fluorescence showed that, whereas the Ca2+ form is well folded, the metal-free form seems strongly disorganized. The Ca2+-induced conformational changes comprise both fast and slow steps, giving thus a structural support for the induction of class II Ca2+-binding sites. The B-repeats may act as rulers or springs that modulate the distance between the interactive A region and the bacterial cell surface.
Josefsson
E
E
Department of Microbiology, Moyne Institute of Preventive Medicine, University of Dublin, Trinity College, Dublin 2, Republic of Ireland. jos.cox@biochem.unie.ch
O'Connell
D
D
Foster
T J
TJ
Durussel
I
I
Cox
J A
JA
D001426:Bacterial Proteins; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002942:Circular Dichroism; D007700:Kinetics; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D020449:Repetitive Sequences, Amino Acid; D017386:Sequence Homology, Amino Acid; D013050:Spectrometry, Fluorescence; D013211:Staphylococcus aureus; D013816:Thermodynamics; D017186:Titrimetry
D001426:Bacterial Proteins; D002135:Calcium-Binding Proteins; D011994:Recombinant Proteins; C115915:SdrD protein, Staphylococcus aureus; D002118:Calcium