ArticleFrontiers in physiology2014
Characterization of heme binding to recombinant α1-microglobulin.
Article in Frontiers in physiology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed, 41 citations in OpenAlex.
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- Update on heme biosynthesis, tissue-specific regulation, heme transport, relation to iron metabolism and cellular energy.Liver international : official journal of the International Association for the Study of the Liver · 2024Review
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- SARS-CoV-2 Proteins Bind to Hemoglobin and Its Metabolites.International journal of molecular sciences · 2021Article
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- Human Plasma and Recombinant Hemopexins: Heme Binding Revisited.International journal of molecular sciences · 2021Article
- Structure, Functions, and Physiological Roles of the Lipocalin αFrontiers in physiology · 2021Review
- Lipocalin Blc is a potential heme-binding protein.FEBS letters · 2021Article
- One ring to bring them all and in the darkness bind them: The trafficking of heme without deliverers.Biochimica et biophysica acta. Molecular cell research · 2021Review
- Evaluation of Peptide/Protein Self-Assembly and Aggregation by Spectroscopic Methods.Molecules (Basel, Switzerland) · 2020Review
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- αFrontiers in physiology · 2020Article
- Expression, Purification and Initial Characterization of Functional αFrontiers in plant science · 2020Article
- Highlighting the Importance of Water Alkalinity Using Phosphate Buffer Diluted With Deionized, Double Distilled and Tap Water, in Lowering Oxidation Effects on Human Hemoglobin Ozonated at High Ozone ConcentrationsFrontiers in molecular biosciences · 2020Article
- Personalizing the safe, appropriate and effective concentration(s) of ozone for a non-diabetic individual and four type II diabetic patients in autohemotherapy through blood hemoglobin analysis.Journal of translational medicine · 2019Article
- rA1M-035, a Physicochemically Improved Human Recombinant αAntioxidants & redox signaling · 2019Article
Corrections and comments
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Authors and funding
6 authors at 2 institutions in 2 countries.
Funding
Abstract
backgroundAlpha-1-microglobulin (A1M), a small lipocalin protein found in plasma and tissues, has been identified as a heme and radical scavenger that may participate in the mitigation of toxicities caused by degradation of hemoglobin. The objective of this work was to investigate heme interactions with A1M in vitro using various analytical techniques and to optimize analytical methodology suitable for rapid evaluation of the ligand binding properties of recombinant A1M versions.
methodsTo examine heme binding properties of A1M we utilized UV/Vis absorption spectroscopy, visible circular dichroism (CD), catalase-like activity, migration shift electrophoresis, and surface plasmon resonance (SPR), which was specifically developed for the assessment of His-tagged A1M.
resultsThe results of this study confirm that A1M is a heme binding protein that can accommodate heme at more than one binding site and/or in coordination with different amino acid residues depending upon heme concentration and ligand-to-protein molar ratio. UV/Vis titration of A1M with heme revealed an unusually large bathochromic shift, up to 38 nm, observed for heme binding to a primary binding site. UV/Vis spectroscopy, visible CD and catalase-like activity suggested that heme is accommodated inside His-tagged (tgA1M) and tagless A1M (ntA1M) in a rather similar fashion although the His-tag is very likely involved into coordination with iron of the heme molecule. SPR data indicated kinetic rate constants and equilibrium binding constants with KD values in a μM range.
conclusionsThis study provided experimental evidence of the A1M heme binding properties by aid of different techniques and suggested an analytical methodology for a rapid evaluation of ligand-binding properties of recombinant A1M versions, also suitable for other His-tagged proteins.
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