Hemoglobin–PEG Interactions Probed by Small-Angle X-ray Scattering: Insights for Crystallization and Diagnostics Applications

Baranova I, Angelova A, Stransky J, Andreasson J, Angelov B
The Journal of Physical Chemistry B (2024 Sep 09)

doi: 10.1021/acs.jpcb.4c03003

Submitted to SASBDB: 2024 May 28 Published in SASBDB: 2024 Sep 10

SASDV62 – Human Haemoglobin (Sigma-Aldrich) in 100mM Sodium Phosphate buffer, pH=7.0, SEC-SAXS Sample: Hemoglobin subunit alpha monomer, 15 kDa Homo sapiens protein Hemoglobin subunit beta monomer, 16 kDa Homo sapiens protein Protoporphyrin IX containing fe monomer, 1 kDa Buffer: 100 mM Sodium Phosphate, pH: 7 Experiment: SAXS data collected at Anton Paar SAXSpoint 2.0, Institute of Biotechnology, Czech Academy of Sciences/Centre of Molecular Structure on 2022 May 24 RgGuinier 2.4 nm Dmax 6.8 nm VolumePorod 105 nm3

SASDV72 – Human Haemoglobin (Sigma-Aldrich) in 100mM Sodium Phosphate buffer, pH=7.0, c=5mg/ml, 1 day after SEC Sample: Hemoglobin subunit alpha monomer, 15 kDa Homo sapiens protein Hemoglobin subunit beta monomer, 16 kDa Homo sapiens protein Protoporphyrin IX containing fe monomer, 1 kDa Buffer: 100 mM Sodium Phosphate, pH: 7 Experiment: SAXS data collected at Anton Paar SAXSpoint 2.0, Institute of Biotechnology, Czech Academy of Sciences/Centre of Molecular Structure on 2022 Sep 30 RgGuinier 2.4 nm Dmax 6.7 nm VolumePorod 103 nm3

SASDV82 – Human Haemoglobin (Sigma-Aldrich) with 5% (w/v) PEG600 in 100mM Sodium Phosphate buffer pH=7.0, c=5 mg/ml Sample: Hemoglobin subunit alpha monomer, 15 kDa Homo sapiens protein Hemoglobin subunit beta monomer, 16 kDa Homo sapiens protein Protoporphyrin IX containing fe monomer, 1 kDa Buffer: 100mM Sodium Phosphate buffer with 5% (w/v) PEG600, pH: 7 Experiment: SAXS data collected at Anton Paar SAXSpoint 2.0, Institute of Biotechnology, Czech Academy of Sciences/Centre of Molecular Structure on 2022 Sep 30 RgGuinier 2.4 nm Dmax 6.7 nm VolumePorod 105 nm3

SASDV92 – Human Haemoglobin (Sigma-Aldrich) with 10% (w/v) PEG600 in 100mM Sodium Phosphate buffer pH=7.0, c=5 mg/ml Sample: Hemoglobin subunit alpha monomer, 15 kDa Homo sapiens protein Hemoglobin subunit beta monomer, 16 kDa Homo sapiens protein Protoporphyrin IX containing fe monomer, 1 kDa Buffer: 100mM Sodium Phosphate buffer with 10% (w/v) PEG600, pH: 7 Experiment: SAXS data collected at Anton Paar SAXSpoint 2.0, Institute of Biotechnology, Czech Academy of Sciences/Centre of Molecular Structure on 2022 Sep 30 RgGuinier 2.4 nm Dmax 7.2 nm VolumePorod 113 nm3

SASDVA2 – Human Haemoglobin (Sigma-Aldrich) with 20% (w/v) PEG600 in 100mM Sodium Phosphate buffer pH=7.0, c=5 mg/ml Sample: Hemoglobin subunit alpha monomer, 15 kDa Homo sapiens protein Hemoglobin subunit beta monomer, 16 kDa Homo sapiens protein Protoporphyrin IX containing fe monomer, 1 kDa Buffer: 100mM Sodium Phosphate buffer with 20% (w/v) PEG600, pH: 7 Experiment: SAXS data collected at Anton Paar SAXSpoint 2.0, Institute of Biotechnology, Czech Academy of Sciences/Centre of Molecular Structure on 2022 Sep 30 RgGuinier 2.6 nm Dmax 7.6 nm VolumePorod 113 nm3

SASDVB2 – Human Haemoglobin (Sigma-Aldrich) in 100mM Sodium Phosphate buffer, pH=7.0, c=5mg/ml, 1 week after SEC Sample: Hemoglobin subunit alpha monomer, 15 kDa Homo sapiens protein Hemoglobin subunit beta monomer, 16 kDa Homo sapiens protein Protoporphyrin IX containing fe monomer, 1 kDa Buffer: 100mM Sodium Phosphate buffer with 5% (w/v) PEG2000, pH: 7 Experiment: SAXS data collected at Anton Paar SAXSpoint 2.0, Institute of Biotechnology, Czech Academy of Sciences/Centre of Molecular Structure on 2022 Oct 6 RgGuinier 2.4 nm Dmax 6.8 nm VolumePorod 107 nm3

SASDVC2 – Human Haemoglobin (Sigma-Aldrich) with 5% PEG2000 in 100mM Sodium Phosphate buffer, pH=7.0, c=5mg/ml Sample: Hemoglobin subunit beta monomer, 16 kDa Homo sapiens protein Hemoglobin subunit alpha monomer, 15 kDa Homo sapiens protein Protoporphyrin IX containing fe monomer, 1 kDa Buffer: 100mM Sodium Phosphate buffer with 5% (w/v) PEG2000, pH: 7 Experiment: SAXS data collected at Anton Paar SAXSpoint 2.0, Institute of Biotechnology, Czech Academy of Sciences/Centre of Molecular Structure on 2022 Oct 6 RgGuinier 2.4 nm Dmax 6.5 nm VolumePorod 92 nm3

SASDVD2 – Human Haemoglobin (Sigma-Aldrich) with 5% (w/v) PEG4000 in 100mM Sodium Phosphate buffer, pH=7.0, c=5mg/ml Sample: Hemoglobin subunit alpha monomer, 15 kDa Homo sapiens protein Hemoglobin subunit beta monomer, 16 kDa Homo sapiens protein Protoporphyrin IX containing fe monomer, 1 kDa Buffer: 100mM Sodium Phosphate buffer with 5% (w/v) PEG4000, pH: 7 Experiment: SAXS data collected at Anton Paar SAXSpoint 2.0, Institute of Biotechnology, Czech Academy of Sciences/Centre of Molecular Structure on 2022 Oct 6 RgGuinier 2.4 nm Dmax 6.1 nm VolumePorod 91 nm3

SASDVE2 – Human Haemoglobin (Sigma-Aldrich) with 10% PEG2000 in 100mM Sodium Phosphate buffer, pH=7.0, c=5mg/ml Sample: Hemoglobin subunit alpha monomer, 15 kDa Homo sapiens protein Hemoglobin subunit beta monomer, 16 kDa Homo sapiens protein Protoporphyrin IX containing fe monomer, 1 kDa Buffer: 100mM Sodium Phosphate buffer with 10% (w/v) PEG2000, pH: 7 Experiment: SAXS data collected at Anton Paar SAXSpoint 2.0, Institute of Biotechnology, Czech Academy of Sciences/Centre of Molecular Structure on 2022 Oct 6 RgGuinier 2.8 nm Dmax 11.3 nm VolumePorod 131 nm3