A highly selective polypeptide binder for human Acetylcholine esterase

T. Tegler, Lotta (författare): Uppsala universitet,Institutionen för biokemi och organisk kemi

Winander, Cecilia (författare): Uppsala universitet,Institutionen för biokemi och organisk kemi

Fromell, Karin (författare): Uppsala universitet,Institutionen för biokemi och organisk kemi

Andersson, Per Ola (författare): Swedish Defence Research Agency, Division of CBRN Defence and Security

Boerjegren, Susanne (författare): Swedish Defence Research Agency, Division of CBRN Defence and Security

Lind, Per (författare): Swedish Defence Research Agency, Division of CBRN Defence and Security

Lundquist, Margaretha (författare): Swedish Defence Research Agency, Division of CBRN Defence and Security

Trogen, Lars (författare): Swedish Defence Research Agency, Division of CBRN Defence and Security

Österlund, Lars (författare): Swedish Defence Research Agency, Division of CBRN Defence and Security

Baltzer, Lars (författare): Uppsala universitet,Institutionen för biokemi och organisk kemi

(creator_code:org_t)

Abstract Ämnesord

Stäng

A highly selective high-affinity polypeptide conjugate binder for human Acetylcholine Esterase (hAChE) has been obtained by coupling a derivative of acridine, a known medium-affinity inhibitor of hAChE, to each member of a 16-membered set of 42-residue polypeptide scaffolds. The best candidate, 4-C10L17-Ac, was identified to have a KD of 10 nM or less in an assay where each polypeptide conjugate was titrated with hAChE in 50 mM phosphate buffer at pH 7.0 and 298K. It was found in a sandwich ELISA to have high selectivity for hAChE in cerebrospinal fluid. Targeting the active site of hAChE by a polypeptide conjugate binder presents a special problem as it is buried deep inside the protein in a cavity that is approximately 20 Å deep. In order to permit simultaneous and cooperative binding of the acridine and the polypeptide to the active site and the AChE surface a fourteen atom spacer was needed. The 9-aminoacridine group was linked to the side chain of a lysine residue in each polypeptide via a spacer prepared from two aminohexanoic acid fragments. The results reinforce the impression that polypeptide conjugates are excellent alternatives to currently used protein binder technologies in diagnostic and therapeutic applications and that the conjugation of enzyme inhibitors to polypeptide scaffolds is a strategy of general applicability in the design of high-affinity binders for enzymes.

Av författaren/redakt...: T. Tegler, Lotta; Winander, Cecili ...; Fromell, Karin; Andersson, Per O ...; Boerjegren, Susa ...; Lind, Per; visa fler...; Lundquist, Marga ...; Trogen, Lars; Österlund, Lars; Baltzer, Lars; visa färre...

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