Characterization of binding and utilization of hemoglobin by <i>Prevotella nigrescens</i>

Guan, Su-Min; Nagata, Hideki; Kuboniwa, Masae; Ikawa, Yoji; Maeda, Kengo; Shizukuishi, Satoshi

doi:10.1034/j.1399-302x.2002.170304.x

“…The strongest binding activity was observed at pH 5.0. Similar pH-dependent hemoglobin-binding activity has also been documented in other periodontal pathogens, such as Porphyromonas gingivalis [16,18], Prevotella nigrescens [19] and Actinobacillus actinomycetemcomitans [20]. Hemoglobin displays conformational changes depending on oxygenation state.…”

Section: Resultssupporting

confidence: 58%

Purification and characterization of a hemoglobin-binding outer membrane protein of Prevotella intermedia

Guan

¹

,

Nagata

²

,

Maeda

³

et al. 2004

FEMS Microbiology Letters

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An outer membrane hemoglobin-binding protein of Prevotella intermedia was identified and purified in the present study. Hemoglobin-binding protein was purified via a series of column chromatographic methods. The molecular mass of the purified protein, which was approximately 60 kDa, was determined by SDS-PAGE. Hemoglobin binding of the protein was examined by Western blot and dot blot assays. Hemoglobin-binding activity was pH dependent; the strongest binding activity occurred at pH 5.0. Globin greatly decreased the binding activity, whereas transferrin, cytochrome C, and catalase exerted no effect. Dissociation constant between the 60-kDa protein and hemoglobin was 1.48 x 10(-8) M as measured by surface plasmon resonance. These results suggest that P. intermedia binds hemoglobin specifically through the 60-kDa outer membrane protein.

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“…The strongest binding activity was observed at pH 5.0. Similar pH‐dependent hemoglobin‐binding activity has also been documented in other periodontal pathogens, such as Porphyromonas gingivalis [16,18], Prevotella nigrescens [19] and Actinobacillus actinomycetemcomitans [20]. Hemoglobin displays conformational changes depending on oxygenation state.…”

Section: Resultssupporting

confidence: 58%

Purification and characterization of a hemoglobin-binding outer membrane protein ofPrevotella intermedia

Guan

¹

,

Nagata

²

,

Maeda

³

et al. 2004

FEMS Microbiology Letters

View full text Add to dashboard Cite

An outer membrane hemoglobin-binding protein of Prevotella intermedia was identified and purified in the present study. Hemoglobin-binding protein was purified via a series of column chromatographic methods. The molecular mass of the purified protein, which was approximately 60 kDa, was determined by SDS-PAGE. Hemoglobin binding of the protein was examined by Western blot and dot blot assays. Hemoglobin-binding activity was pH dependent; the strongest binding activity occurred at pH 5.0. Globin greatly decreased the binding activity, whereas transferrin, cytochrome C, and catalase exerted no effect. Dissociation constant between the 60-kDa protein and hemoglobin was 1.48 x 10(-8) M as measured by surface plasmon resonance. These results suggest that P. intermedia binds hemoglobin specifically through the 60-kDa outer membrane protein.

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“…According a report of HbBP of P. nigrescens [22], it was also active in low pH buffers than at higher pH. Its activity was resistant to heating at 60°C for 10 min, but obvious decrease was observed heating at 80°C.…”

Section: Discussionmentioning

confidence: 98%

Hemoglobin binding activity and hemoglobin-binding protein of prevotella nigrescens

Miyashita

¹

,

Oishi

²

,

Kiso

³

et al. 2010

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Prevotella nigrescens, lacking siderophores was found to bind to the hemoproteins. The binding was observed also in the envelope which was prepared by sonication of the cell. The binding occurred in the pH-dependent manner; the binding was observed below neutral pHs of the incubation mixtures but only slightly observed in the neutral and alkaline pHs. Furthermore, hemoglobin bound to the envelope was dissociated at high pHs buffers. Maximum amounts of hemoglobin bound to 1 mg envelope was 51.2 μg. Kd for the reaction at pH 5.0 was 2.1 × 10-10M (210 pM). From the dot blot assay, hemoglobin could bind to a protein solubilized from the envelope by a detergent, referred to as hemoglobin-binding protein (HbBP), then it was purified by the sequential procedures of ion exchange chromatography, affinity chromatography and isoelectric focusing. Molecular weight and isoelectric point of the HbBP were 46 kDa and 6.1, respectively.

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Characterization of binding and utilization of hemoglobin by Prevotella nigrescens

Cited by 6 publications

References 35 publications

Purification and characterization of a hemoglobin-binding outer membrane protein of Prevotella intermedia

Purification and characterization of a hemoglobin-binding outer membrane protein of Prevotella intermedia

Purification and characterization of a hemoglobin-binding outer membrane protein ofPrevotella intermedia

Hemoglobin binding activity and hemoglobin-binding protein of prevotella nigrescens

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