Single-site cys-substituting mutation of human lectin galectin-2: Modulating solubility in recombinant production, reducing long-term aggregation, and enabling site-specific monoPEGylation

Wang, H., He, L., Lensch, M., Gabius, H. J., Fee, C. J. and Middelberg, A. P.J. (2008) Single-site cys-substituting mutation of human lectin galectin-2: Modulating solubility in recombinant production, reducing long-term aggregation, and enabling site-specific monoPEGylation. Biomacromolecules, 9 11: 3223-3230. doi:10.1021/bm800801b


Author Wang, H.
He, L.
Lensch, M.
Gabius, H. J.
Fee, C. J.
Middelberg, A. P.J.
Title Single-site cys-substituting mutation of human lectin galectin-2: Modulating solubility in recombinant production, reducing long-term aggregation, and enabling site-specific monoPEGylation
Journal name Biomacromolecules   Check publisher's open access policy
ISSN 1525-7797
Publication date 2008-11-01
Year available 2008
Sub-type Article (original research)
DOI 10.1021/bm800801b
Open Access Status
Volume 9
Issue 11
Start page 3223
End page 3230
Total pages 8
Editor Ann-Christine Albertsson
Place of publication U.S.
Publisher American Chemical Society
Language eng
Subject C1
929999 Health not elsewhere classified
119999 Medical and Health Sciences not elsewhere classified
Abstract The effector capacity of endogenous lectins on cell adhesion/growth prompts studies to turn them into pharmaceutically stable forms. Using human galectin-2 as a proof-of-principle model, we first introduced mutations at the site of one of the two Cys residues, that is, C57A, C57M, and C57S. Only the C57M variant was expressed in bacteria in soluble form in high yield. No notable aggregation of the modified homodimeric lectin occurred during 3 weeks of storage. This mutational process also facilitated the site-directed introduction of poly(ethylene glycol) into the remaining sulfhydryl group (Cys75). Product analysis revealed rather complete conjugation with one chain per subunit in the homodimer. We note that neither the secondary structure alteration nor the absence of binding ability to a glycoprotein (asialofetuin) was observed. The results thus document the feasibility of tailoring a human galectin for enhanced stability to aggregation as well as monoPEGylation, which enables further testing of biological properties including functionality as growth regulator and the rate of serum clearance.
Q-Index Code C1
Q-Index Status Confirmed Code

Document type: Journal Article
Sub-type: Article (original research)
Collection: 2009 Higher Education Research Data Collection
 
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Created: Sat, 18 Apr 2009, 00:54:03 EST by Vicki Thompson on behalf of School of Chemical Engineering