Journal Article

Polyionic fusion peptides function as specific dimerization motifs

Susanne A. Richter, Kay Stubenrauch, Hauke Lilie and Rainer Rudolph

in Protein Engineering, Design and Selection

Volume 14, issue 10, pages 775-783
Published in print October 2001 | ISSN: 1741-0126
Published online October 2001 | e-ISSN: 1741-0134 | DOI: https://dx.doi.org/10.1093/protein/14.10.775
Polyionic fusion peptides function as specific dimerization motifs

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The de novo design of a molecular adapter for directed association and covalent linkage of two polypeptides is presented. Using peptides containing charged amino acid residues and an additional cysteine residue (AlaCysLys8 and AlaCysGlu8) we demonstrate that the electrostatic interaction promotes the association of two synthetic peptides and, subsequently, disulfide bond formation. The reaction depends on both the redox potential and on the ionic strength of the buffer. Varying the redox potential, the interaction of the peptides was quantified by a ΔG0′ of 6.6 ± 0.2 kcal/mol. Heterodimerization of the peptides is highly specific, a competition of association by other cysteine containing compounds could not be observed. Two proteins comprising cysteine-containing polyionic fusion peptides, a modified Fab fragment and an α-glucosidase fusion, could be specifically conjugated by directed association and subsequent disulfide bond formation. Both proteins retain their functional characteristics within the bifunctional conjugate: enzymatic activity of the αglucosidase and antigen-binding capacity of the Fab fragment are equivalent to the non-conjugated components.

Keywords: directed association; disulfide bond; heterodimerization; polyarginine; polyglutamic acid

Journal Article.  7104 words.  Illustrated.

Subjects: Proteins

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