Transfer RNA binding to human serum albumin: a model for protein-RNA interaction.
Protein-RNA complexation is essential in cell biological functions. Transfer RNAs are bound to aminoacyl-tRNA synthetases for the translation of the genetic code during protein synthesis, while ribonucleoproteins bind RNA in posttranscriptional regulation of gene expression. A recent report showed the interacton of human serum albumin (HSA) with DNA duplex, in which two binding sites with strong and weak association constants were detected. We now examine the interaction of tRNA with human serum albumin (HSA) in aqueous solution at physiological conditions, using a constant RNA concentration of 12.5 mM (phosphate) and various HSA contents of 0.04 to 0.6 mM. Affinity capillary electrophoresis and FTIR spectroscopic methods were used to determine the protein binding mode, the association constant, sequence preference, and the biopolymer secondary structural changes in the HSA-RNA complexes. Spectroscopic evidence showed two types of HSA-RNA complexes with an overall binding constant of K = 1.45 x 10(4) M(-1). The major binding sites were located on the G-C bases and the backbone PO2 group. The protein-RNA interaction stabilizes the HSA secondary structure, and no major alterations of A-RNA structure or protein conformation occurred.
Malonga
Herman
H
Department of Chemistry-Biology, University of Québec at Trois-Rivières, Québec, Canada.
Neault
Jean-Francois
JF
Tajmir-Riahi
Heidar-Ali
HA
D001665:Binding Sites; D019075:Electrophoresis, Capillary; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D046911:Macromolecular Substances; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D012331:RNA, Fungal / Q000737:chemistry / Q000378:metabolism; D012343:RNA, Transfer / Q000737:chemistry / Q000378:metabolism*; D012709:Serum Albumin / Q000737:chemistry / Q000378:metabolism*; D017550:Spectroscopy, Fourier Transform Infrared
D046911:Macromolecular Substances; D012331:RNA, Fungal; D012709:Serum Albumin; D012343:RNA, Transfer