THE DEVELOPMENT AND APPLICATION OF CLICKABLE LIPID ANALOGS FOR ELUCIDATING AND HARNESSING LIPID FUNCTIONS - Click Chemistry in Glycoscience: New Developments and Strategies - page 96

formation when compared to tris-(2-carboxyethyl)phosphine (TCEP). Subsequently,
these compounds were exploited to characterize covalent protein adducts from RKO
cell lines using biotinylation via either CuAAC or the Staudinger ligation, followed by
in-gel and mass spectrometric analysis. In these studies, the authors noted problems
eluting biotin-conjugated proteins from beads when using the Staudinger ligation,
and thus derivatization via CuAAC was found to be more effective. Also, alkynyl-
HNE 43b was observed to yield less nonspecific labeling compared to azido-HNE
43a . Proteomic experiments resulted in the labeling of a number of proteins involved
in stress responses, such as heat shock proteins.
4.7 CONCLUSION
From the examples described in this chapter, it is clear that, following the initial
formulation of the philosophy of click chemistry, this strategy has quickly led to
significant impacts regarding the study and application of lipid structures. In par-
ticular, the bioorthogonal nature of these recently discovered reactions has allowed
researchers to infiltrate the highly complex environments of cellular membranes for
efforts aimed at further understanding and harnessing the activities of target lipids.
From the direct labeling and imaging of specific lipid and membrane structures in
living samples to the ability to fish out proteins that are modified through interac-
tions with lipids, studies facilitated by click chemistry show great promise for a
paradigm shift in the methods by which these enigmatic molecules are analyzed.
However, many of the studies discussed herein represent the initial wave of reports
that showcase the efficacy of click-chemistry-based studies. In the future, the suc-
cess of these approaches will be judged by the extent to which they are successfully
implemented to answer important questions pertaining to the activity and properties
of lipids, as well as the applications that they enable for exploiting these proper-
ties. Nevertheless, the early reports point to a bright future for click-chemistry-based
lipid studies.
REFERENCES
1. Cho, W. H.; Stahelin, R. V. Ann. Rev. Biophys. Biomol. Struct. 2005 , 34 , 119.
2. Lemmon, M. A. Nat. Rev. Mol. Cell. Biol. 2008 , 9 , 99.
3. Resh,M.D. Nat. Chem. Biol. 2006 , 2 , 584.
4. Jahn, R.; Lang, T.; Sudhof, T. C. Cell. 2003 , 112 , 519.
5. Brochu, H.; Polidori, A.; Pucci, B.; Vermette, P. Curr. Drug. Deliv. 2004 , 1 , 299.
6. Ferrara, K. W.; Borden, M. A.; Zhang, H. Acc. Chem. Res. 2009 , 42 , 881.
7. Mintzer, M. A.; Simanek, E. E. Chem. Rev. 2009 , 109 , 259.
8. Vermette, P.; Meagher, L.; Gagnon, E.; Griesser, H. J.; Doillon, C. J. J. Cont. Rel. 2002 ,
80 , 179.
9. Cho, W. W.; Bittova, L.; Stahelin, R. V. Anal. Biochem. 2001 , 296 , 153.