DESIGN AND SYNTHESIS OF A NOVEL BIOTINYLATED PHOTOREACTIVE INSULIN FOR RECEPTOR ANALYSIS

被引：9

作者：

FABRY, M ^{[1
]}

BRANDENBURG, D ^{[1
]}

机构：

[1] RHEIN WESTFAL TH AACHEN,DEUTSCH WOLLFORSCHUNGSINST,VELTMANPL 8,W-5100 AACHEN,GERMANY

来源：

BIOLOGICAL CHEMISTRY HOPPE-SEYLER | 1992年 / 373卷 / 03期

关键词：

INSULIN; SEMISYNTHESIS; INSULIN RECEPTOR; PHOTOAFFINITY LABELING;

D O I：

10.1515/bchm3.1992.373.1.143

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

B1-(4-Azido-salicyloyl)-[B1-biocytin,B2-lysine]insulin was synthesized by double Edman degradation of A1,B29-Msc2-insulin and stepwise acylation at the N-terminus of the B-chain. This derivative is homogeneous in RP-HPLC and has a biological in vitro activity of 20% and receptor binding of 15%, relative to insulin. Radioiodination and HPLC gave the B1-labelled I-125-derivative (I) as well as the 4 isomers with I-125-labelled tyrosine (A14, A19, B16, B26). UV-induced crosslinking of I with insulin receptors led to specific labelling of the alpha-subunit (M(r) 130 000). The peptide bond Lys(B2)-Asp(B3) is completely cleavable by trypsin (EC 3.4.21.4). I is thus a new tool for the analysis of the hormone-binding region by making possible the isolation of tryptic, biotinylated receptor fragments labelled by the dipeptide I-125-4-azidosalicyloyl-biocytinyl-Lys.

引用

页码：143 / 150

页数：8

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