Sortase-catalyzed transformations that improve the properties of cytokines.
Proc Natl Acad Sci U S A
; 108(8): 3169-74, 2011 Feb 22.
Article
em En
| MEDLINE
| ID: mdl-21297034
ABSTRACT
Recombinant protein therapeutics often suffer from short circulating half-life and poor stability, necessitating multiple injections and resulting in limited shelf-life. Conjugation to polyethylene glycol chains (PEG) extends the circulatory half-life of many proteins, but the methods for attachment often lack specificity, resulting in loss of biological activity. Using four-helix bundle cytokines as an example, we present a general platform that uses sortase-mediated transpeptidation to facilitate site-specific attachment of PEG to extend cytokine half-life with full retention of biological activity. Covalently joining the N and C termini of proteins to obtain circular polypeptides, again executed using sortase, increases thermal stability. We combined both PEGylation and circularization by exploiting two distinct sortase enzymes and the use of a molecular suture that allows both site-specific PEGylation and covalent closure. The method developed is general, uses a set of easily accessible reagents, and should be applicable to a wide variety of proteins, provided that their termini are not involved in receptor binding or function.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Polietilenoglicóis
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Proteínas de Bactérias
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Cisteína Endopeptidases
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Aminoaciltransferases
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Estabilidade Proteica
Limite:
Animals
Idioma:
En
Ano de publicação:
2011
Tipo de documento:
Article