Labeling of Peptides

Segment in model Hydro- Negatively charged residues Hydrophobic labelling of peptide Tryptic limit peptide... 

Galardy, R.E., Craig, L.C., Jamieson, J.D., and Printz, M.P. (1974) Photoaffinity labeling of peptide hormone binding sites./. Biol. Chem. 249, 3510-3518. 

Here, we discuss a solid-state 19F-NMR approach that has been developed for structural studies of MAPs in lipid bilayers, and how this can be translated to measurements in native biomembranes. We review the essentials of the methodology and discuss key objectives in the practice of 19F-labelling of peptides. Furthermore, the preparation of macroscopically oriented biomembranes on solid supports is discussed in the context of other membrane models. Two native biomembrane systems are presented as examples human erythrocyte ghosts as representatives of eukaryotic cell membranes, and protoplasts from Micrococcus luteus as membranes... 

B. de Bruin, B. Kuhnast, F. Hinnen, L. Yaouancq, M. Amessou, L. Johannes, A. Samson, R. Boisgard, B. Tavitian, F. Dolle, 1-[3-(2-[ F]Fluoropyridin-3-yloxy)propyl]pyrrole-2,5-dione Design, synthesis and radiosynthesis of a new [ F]fluoropyridine-based maleimide reagent for the labeling of peptides and proteins, Bioconj. Chem. 16 (2005) 406-420. 

B. Kuhnast, F. Dolle, B. Tavitian, Fluorine-18 labeling of peptide nucleic acids, J. Label. Compds Radiopharm. 45 (2001) 1-11. 

Fig. 8. 750 MHz H-I5N HMQC (A) and H NOESY (B) HRMAS NMR spectra of Gly-Pro-Gly-Arg-Ala-Phe on TantaGel resin swollen with DMSO-

N and 13C labeling of peptides facilitates the study of their molecular dynamics in solution by measurements of relaxation parameters (42,43). Heteronuclear relaxation times and heteronuclear NOEs are predominantly affected by the dipole-dipole interaction of the heteronucleus with the directly attached proton. Since the intemuclear (i.e., chemical bonding) distances are known from the molecular geometry, correlation times for overall and internal motions can be determined. 

M. Munchbach, M. Quadroni, G. Miotto, and P. James, Quantitation and facilitated de novo sequencing of proteins by isotopic N-terminal labeling of peptides with a fragmentation-directing moiety, Anal. Chem. 72 (2000), 4047-4057. 

Alberto R, Pak JK, van Staveren D, Mundwiler S, Benny P. Mono-, bi-, or tridentate ligands The labeling of peptides with 99mTc-carbonyls. Biopolymers 2004 76 324-333. 

A second approach relies on the traceless Staudinger ligation between an appropriately functionalized phosphine and an azide to produce a new amide bond [54]. This method has found extensive utility in the labeling of peptides and proteins [215]. Importantly, this method has also been used for the construction of native iV-linked glycans through the reaction between a glycosyl azide and a phosphine containing amino acid [216,217]. 

There are a number of alternative ways to achieve isotopic labelling of peptides for quantitative purposes. One can start at the protein level, like in ICAT ... 

D.R. Goodlett, A. Keller, J.D. Watts, R. Newitt, E.C. Yi, S. Purvine, J.K. Eng, P. von Haller, R. Aebersold, E. Kolker, Differential stable isotope labeling of peptides for quantitation and de novo sequence derivatization. Rapid Commun. Mass Spectrom., 15 (2001) 1214. 

Freedlender and Yalow and Berson, have discussed in detail the labeling of peptide hormones for radioimmunoassay (F8, Yl, Y2). Readers are referred to their papers for further information on this subject. 

GREENLAND, W.E., BLOWER, P.J., Water-soluble phosphines for direct labelling of peptides with technetium and rhenium Insights from electrospray mass spectrometry, Bioconjug. Chem. 16 (2005) 939-948. 

The labeling of peptides with Tc-99m-carbonyls. Biopolymers 76 324-333 Alves S, Paulo A, Correia JDG, Gano L, Smith CJ, Hoffman TJ, Santos I (2005) Pyrazolyl derivatives as bifunctional chelators for labeling tiunom-seeking peptides with the/ac-M(CO)3 moiety (M = Tc-99m, Re) synthesis, characterization, and biological behavior. Bioconjugate Chem 16 438-449... 

Bolton-Hunter Reagent Although not the first to be synthesized, the reagent developed by Bolton and Hunter (Bolton and Hunter, 1973), A -succinimidyl-3-(4-hydroxyphenyl)propionate (Figure 76.6), has found prominent use for the labeling of peptides and small molecules (biotin, melatonin, clonazepam, and so on). 

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