Interaction of bilirubin with native and protein-depleted human erythrocyte membranes.
In order to study the role of membrane proteins in bilirubin (BR) binding phenomenon, selective removal of membrane proteins was carried out using various reagents, namely, ethylenediamine tetraacetic acid (EDTA), sodium hydroxide (NaOH), 3,5-diiodosalicylic acid, lithium salt (LIS), dimethylmaleic anhydride (DMMA), sodium iodide (NaI), o-phenanthroline-cupric sulfate (CuP) and phenanthroline-cupric sulfate containing 2-mercaptoethanol (CuP-mercaptoethanol). Effects of these treatments on the conformation and BR binding properties of the membrane were studied using circular dichroism (CD) spectroscopy as well as estimation of membrane-bound BR by diazotised-color reaction. Though a significant amount of protein (ranging from 23-69%) was lost from the membranes upon these treatments, only a small decrease (3-13%) was observed in BR binding, being maximum with NaOH-treated membranes. However, DMMA and NaI treatments produced a little increase in BR binding. Conformation of the membrane was retained to a significant extent as indicated by far-UV CD spectra upon these treatments except in DMMA and NaI treatments which resulted in the perturbation in CD spectra. Taken together, these results suggest that membrane proteins play little role in BR binding, rather act as barriers in BR binding phenomenon.
Rashid
Huma
H
Interdisciplinary Biotechnology Unit, Aligarh Muslim University, Aligarh, India.
Tayyab
Saad
S
D001663:Bilirubin / Q000378:metabolism*; D002942:Circular Dichroism; D019327:Copper Sulfate / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D007202:Indicators and Reagents; D008094:Lithium / Q000494:pharmacology; D008299:Maleic Anhydrides / Q000494:pharmacology; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism; D011485:Protein Binding; D012972:Sodium Hydroxide / Q000494:pharmacology; D012974:Sodium Iodide / Q000494:pharmacology
D007202:Indicators and Reagents; D008299:Maleic Anhydrides; D008565:Membrane Proteins; D012972:Sodium Hydroxide; C007474:2,3-dimethylmaleic anhydride; D008094:Lithium; D004492:Edetic Acid; D012974:Sodium Iodide; D019327:Copper Sulfate; D001663:Bilirubin