Dataset Information

Structural Stability of Streptococcal Serum Opacity Factor.

ABSTRACT: Streptococcal serum opacity factor (SOF) is a protein that clouds the plasma of multiple mammalian species by disrupting high density lipoprotein (HDL) structure. Intravenous infusion of low dose SOF (4 µg) into mice reduces their plasma cholesterol concentrations?~?40% in 3 h. Here we investigated the effects of pH, ionic strength, temperature, and denaturation with guanidinium chloride (GdmCl) on SOF stability and its reaction vs HDL. SOF stability was tested by pre-incubation of SOF at various temperatures, pH's, and GdmCl concentrations and measuring the SOF reaction rate at pH 7.4 and 37?°C. SOF retained activity at temperatures up to 58?°C, at pH 4 to 10, and in 8.5 M GdmCl after being returned to standard buffer conditions. The effects of GdmCl, pH, and ionic strength on the SOF reaction rates were also measured. SOF was inactive at GdmCl???1 M; SOF was most active at pH 5, near its isoelectric point and at an ionic strength of 3 (in NaCl). These data reveal that SOF is a stable protein and suggest that its activity is determined, in part, by the effects of pH and ionic strength on its overall charge relative to that of its reaction target, HDL.

SUBMITTER: Yelamanchili D

PROVIDER: S-EPMC5448415 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature

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Structural Stability of Streptococcal Serum Opacity Factor.

Yelamanchili Dedipya D Gillard Baiba K BK Gotto Antonio M AM Pownall Henry J HJ Rosales Corina C

The protein journal 20170601 3

Streptococcal serum opacity factor (SOF) is a protein that clouds the plasma of multiple mammalian species by disrupting high density lipoprotein (HDL) structure. Intravenous infusion of low dose SOF (4 µg) into mice reduces their plasma cholesterol concentrations ~ 40% in 3 h. Here we investigated the effects of pH, ionic strength, temperature, and denaturation with guanidinium chloride (GdmCl) on SOF stability and its reaction vs HDL. SOF stability was tested by pre-incubation of SOF at variou ...[more]

PMID: 28374173

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Project description:The group A Streptococcus (GAS) sof gene encodes the serum opacity factor protein, which is capable of opacifying mammalian sera and binding at least two host proteins, fibronectin and fibrinogen. The sof gene exists in approximately 50% of clinical isolates, and there is a classical association of so-called nephritogenic strains with the opacity factor-positive phenotype. In both a type emm49 strain and a type emm12 strain, the sequences upstream of the 5' end of sof and downstream of the putative terminator were determined to be nearly identical to a region in the M type 1 genome approximately 10 kb upstream of the emm1 gene. This close genetic linkage is likely reflected in the strict correlation of opacity factor phenotype with specific emm genotypes. A new fibronectin-binding protein gene, sfbX, was discovered immediately downstream of sof in emm12 and emm49 strains and in several other sof-positive strains. The sof and sfbX genes were found to be expressed on the same transcription unit, which was correlated with the putative promoter and rho-independant terminator sequences that flank these two genes. The sfbX genes from different emm types are predicted to encode approximately 650-residue surface-bound proteins sharing 89 to 92% sequence identity. SfbX residues approximately 1 to 480 are not highly similar to those of other known proteins, with the closest match being the Staphylococcus aureus coagulase protein. The remaining portions of these proteins (residues 481 to 650) contain four putative fibronectin-binding repeats highly similar to those of other streptococcal fibronectin-binding proteins and a potential LP(X)SG cell wall anchor motif. Targeted in-frame allelic-exchange mutagenesis, complementation, and heterologous-expression studies found that serum opacification is encoded by sof alone and that sfbX encodes a fibronectin-binding function. A recombinant SfbX protein was found to bind immobilized fibronectin and to partially inhibit GAS adherence to fibronectin. The sfbX gene was found to be present only in sof-positive strains, and together these genes could influence the spectrum of tissues colonized by sof-positive GAS.

Dataset Information

Structural Stability of Streptococcal Serum Opacity Factor.

Publications

Structural Stability of Streptococcal Serum Opacity Factor.

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