Transglutaminase-mediated methods for site-selective modification of human growth hormone.
Two methods for the site-selective modification of native human growth hormone (hGH) using microbial transglutaminase were developed. In the first method, 1,3-bisaminoxypropane was attached to hGH, providing a direct incorporation of reactive aminoxy groups for further modification. The reaction was shown to be selective for Gln(141), with minor modification at Gln(40). In the second method, modified glutamine substrates were developed for attachment to Lys(145) in hGH. A series of glutamine-substrates were screened, and it was shown that microbial transglutaminase was selective towards substitutions on the glutamine core structure. Products from both methods could be transformed to site selectively mono-PEGylated hGH-derivatives in good isolated yield.
Buchardt
Jens
J
Protein Technology, Novo Nordisk A/S, Måløv, Denmark. jbuc@novonordisk.com
Selvig
Helle
H
Nielsen
Per Franklin
PF
Johansen
Nils Langeland
NL
D001665:Binding Sites; D002384:Catalysis; D019382:Human Growth Hormone / Q000737:chemistry*; D006801:Humans; D015394:Molecular Structure; D011503:Transglutaminases / Q000737:chemistry* / Q000378:metabolism
D019382:Human Growth Hormone; D011503:Transglutaminases