N-乙酰苯丙氨酰基[s4U]苯丙氨酸转运核糖核酸在核糖体P位点与蛋白质S10的交联。

Crosslinking of N-acetyl-phenylalanyl [s4U]tRNAPhe to protein S10 in the ribosomal P site.

作者信息

Riehl N, Remy P, Ebel J P, Ehresmann B

出版信息

Eur J Biochem. 1982 Nov 15;128(2-3):427-33. doi: 10.1111/j.1432-1033.1982.tb06982.x.

DOI:10.1111/j.1432-1033.1982.tb06982.x

Abstract

In order to identify ribosomal components involved in the peptidyl-tRNA binding site on the ribosome, tRNAPhe molecules were prepared in which cytidine residues had been chemically converted into 4-thiouridine (S4U). This nucleoside is photoactive at 335 nm and able to form covalent bonds with nearby nucleophilic groups. The thiolated AcPhe-tRNAPhe was bound to the ribosomal P site in the presence of poly(U) as verified by puromycin reactivity. Direct irradiation of the AcPhe-[s4U]tRNAPhe poly(U) 70-S ribosome complex induced crosslinking of the tRNA molecule exclusively to 30-S subunits. Analysis of the covalent complex revealed that AcPhe-[s4U]tRNAPhe was specifically crosslinked to protein S10.

摘要

为了鉴定核糖体上肽基 - tRNA结合位点所涉及的核糖体组分，制备了其中胞苷残基已被化学转化为4 - 硫尿苷（S4U）的苯丙氨酸tRNA（tRNAPhe）分子。这种核苷在335nm处具有光活性，并且能够与附近的亲核基团形成共价键。如通过嘌呤霉素反应性所证实的，在聚尿苷酸（poly(U)）存在下，硫醇化的乙酰苯丙氨酰 - tRNAPhe（AcPhe - tRNAPhe）与核糖体P位点结合。对乙酰苯丙氨酰 - [s4U]tRNAPhe - poly(U) - 70 - S核糖体复合物的直接照射仅诱导了tRNA分子与30 - S亚基的交联。对共价复合物的分析表明，乙酰苯丙氨酰 - [s4U]tRNAPhe与蛋白质S10特异性交联。