In-vivo labeling

In vivo labeling of sea urchin eggs revealed that fertilization results in an eightfold increase in 32P orthophosphate incorporation into tyrosine residues of egg proteins because of an increase in the activity of plasma membrane-bound tyrosine protein kinases (Ribot et al., 1984). Tyrosine phosphorylation is therefore believed to be an important step in the events following fertilization. 

The use of the in vivo labeling methods described above is limited by the fact that the sample must be grown in the presence of the labeling isotopes. In many cases, it is not feasible to perform in vivo metabolic labeling. For example, for human clinical samples it is not possible to perform in vivo labeling and yet it is highly desirable to obtain accurate quantitative information on protein expression levels within these samples. Therefore, robust methods are needed for quantitation of protein levels in the absence of in vivo labeling with isotopes. 

The results obtained with the ICAT labeling strategy are similar, in principle, to those obtained by in vivo labeling with stable isotopes in that the relative ratios of proteins from different samples are obtained (Fig. 3.2). The important difference between the methods is that the ICAT-labeling procedure is performed in vitro on protein lysates. Therefore, the ICAT-labeling strategy can be applied to samples that cannot be labeled in vivo. Because the ICAT-labeling method can be used with virtually any sample, it... 

The strategy described has been demonstrated by the in vivo labeling of proteins in both bacterial and mammalian systems thereby making it potentially useful for future bioimaging and proteomics applications [163],... 

Paganelli, G., Riva, P., Deleide, G., Clivio, A., Chiolerio, F., Scassellati, G.A., Malcovati, M., and Siccardi, A.G. (1988) In vivo labeling of biotinylated monoclonal antibodies by radioactive avidin A strategy to increase tumor radiolocalization. Int.. Cancer 2, 121-125. 

Keywords Amplification Fluorescent reporter Fluorophore FRET In vitro In vivo Labeling Lanthanide chelate Multiplexing Nanoparticle Quantum dot Transition metal complex... 

K. V. Investigation of the early steps of molybdopterin biosynthesis in Escherichia coli through the use of in vivo labelling studies, J. Biol. Chem. 

Lu E, Wagner WR, Schellenberger U, Abraham JA, Klibanov AL, WouRe SR et al. Targeted in vivo labeling of receptors for vascular endothelial growth factor approach to identification of ischemic tissue. Circulation 2003 108 97-103... 

Fitzgerald et al,54 studied the monitoring of calcium metabolism in patients in the final stages of renal disease using serum by AMS after isotopic labelling with a 41 Ca radiotracer. The authors hypothesized that bone resorption can be studied directly by serial measurements of the 41Ca/Ca,otal ratio in serum after in vivo labelling of the calcium pool with 41 Ca.54... 

Schutte, B., Reynders, M. M. J., Bosman, F. T., and Blijham, G. H. (1987) Studies with antibromodeoxyuridine antibodies 11 simultaneous immunocytochemical detection of antigen expression and DNA synthesis by in vivo labelling of intestinal mucosa. J. Histochem. Cytochem. 35,371—374. 

Laporte A-M, Lima L, Gozlan H, Hamon M. Selective in vivo labelling of brain 5-HT1A receptors by [3H]WAY 100635 in the mouse. Eur J Pharmacol 1994 27 505-514. 

This probe was found to target HDACs in proteomes, and was also capable of in vivo labeling. In addition, several HD AC-associated proteins were also labeled by SAHA-BPyne, indicating that these proteins are in close proximity to HDAC active sites and may regulate substrate recognition and activity [90]. For a more detailed survey of the appbcations of photoaffinity labeling as it applies to ABPP, the reader is directed to a review by Overkleeft et al. [91]... 

Staub I, Sieber SA (2008) P-Lactams as selective chemical probes for the in vivo labelling of bacterial enzymes involved in cell wall biosynthesis, antibiotic resistance, and virulence. J Am Chem Soc 130 13400-13409... 

This bioorthogonality was exploited to achieve in vivo labelling in both E. coli overexpressing LasR-LBD and P. aeruginosa. The bacteria were fed probe for 12 h before washing to remove unused lactone. The cell-permeable BODIPY-oxime was then fed to the cells for a further 12 h at pH 6.6 at 8 °C, after which the bacteria... 

Molina, H., Parmigiani, G., Pandey, A. (2005). Assessing reproducibility of a protein dynamics study using in vivo labeling and liquid chromatography tandem mass spectrometry. Anal. Chem. 77, 2739-2744. 

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