Isopept day

The isopeptag is a protein tag of sixteen amino acids that can be used to determine protein-protein interactions .

properties

By coupling the isopeptide tag to the binding partner of the recombinant protein, the binding is fixed by crosslinking via an isopeptide bond on the isopeptide tag (TDKDMTITFTNKKDAE). The isopeptag is derived from the pilin protein Spy0128 from Streptococcus pyogenes . Spy0128 contains two intramolecular, autocatalytically formed isopeptide bonds that cause the stability of a pilus . By removing a β-sheet of the Spy0128 and thus one of the two isopeptide bonds, the isopeptide tag was created. The isopeptide bond takes place at temperatures between 4 ° C and 37 ° C, at pH values between five and eight, regardless of redox and in the presence of common surfactants . The enzymatic isopeptag can also be divided into the isopeptag and an enzymatically active protein fragment, which - coupled to two different proteins - can mediate a Förster resonance energy transfer between two fluorophores .

Individual evidence

↑ ^a ^b B. Zakeri, M. Howarth: Spontaneous intermolecular amide bond formation between side chains for irreversible peptide targeting. In: Journal of the American Chemical Society . Volume 132, Number 13, April 2010, ISSN 1520-5126 , pp. 4526-4527, doi : 10.1021 / ja910795a , PMID 20235501 .
^ HJ Kang, F. Coulibaly, F. Clow, T. Proft, EN Baker: Stabilizing isopeptide bonds revealed in gram-positive bacterial pilus structure. In: Science . Volume 318, number 5856, December 2007, ISSN 1095-9203 , pp. 1625-1628, doi : 10.1126 / science.1145806 , PMID 18063798 .
↑ H. Abe, R. Wakabayashi, H. Yonemura, S. Yamada, M. Goto, N. Kamiya: Split Spy0128 as a potent scaffold for protein cross-linking and immobilization. In: Bioconjugate Chemistry . Volume 24, Number 2, February 2013, ISSN 1520-4812 , pp. 242-250, doi : 10.1021 / bc300606b , PMID 23350748 .

[zakeri-1] B. Zakeri, M. Howarth: Spontaneous intermolecular amide bond formation between side chains for irreversible peptide targeting. In: Journal of the American Chemical Society . Volume 132, Number 13, April 2010, ISSN 1520-5126 , pp. 4526-4527, doi : 10.1021 / ja910795a , PMID 20235501 .

[2] HJ Kang, F. Coulibaly, F. Clow, T. Proft, EN Baker: Stabilizing isopeptide bonds revealed in gram-positive bacterial pilus structure. In: Science . Volume 318, number 5856, December 2007, ISSN 1095-9203 , pp. 1625-1628, doi : 10.1126 / science.1145806 , PMID 18063798 .

[3] H. Abe, R. Wakabayashi, H. Yonemura, S. Yamada, M. Goto, N. Kamiya: Split Spy0128 as a potent scaffold for protein cross-linking and immobilization. In: Bioconjugate Chemistry . Volume 24, Number 2, February 2013, ISSN 1520-4812 , pp. 242-250, doi : 10.1021 / bc300606b , PMID 23350748 .