Protein localization

The activation of NFAT is regulated by calcium. In resting cells, NFAT proteins localize in the cytoplasm... 

Johnson E.W., Eller P. and Jafek B.W. (1993). An immunoelectron microscopic comparison of olfactory marker protein localization in the supranuclear regions of the rat olfactory epithelium and vomeronasal organ neuroepithelium. Acta Oto-Laryngol 113, 766-771. 

Downstream events protein localization spindle orientation cell fate resolution... 

To see whether similar defects in asymmetric cell divisions and protein localization can be seen in other cdc2 mutant combinations, we used a stock... 

Chia If it is just a timing issue we wouldn t expect the asymmetrical protein localization to be affected. 

Horton, P., Park, K. J., Obayashi, T. et al. 2007. WoLF PSORT Protein localization predictor, Nucleic Acids Res., 35(Web Server issue) W585-W587. 

The identity of TES-32 and CTL-1 was confirmed by polyclonal antibodies to recombinant CTL-1, which bound to native TES-32, and by monoclonal antibody Tcn-3, raised to native TES-32 (Maizels et al, 1987), which specifically recognized recombinant CTL-1. The CTL-1 sequence also contained three sites for Afglycosylation, which had previously been shown to be present on TES-32 (Page and Maizels, 1992). Both Tcn-3 and polyclonal antibody to the recombinant CTL-1 protein localize to the cuticle of the infective larvae by immunoelectron microscopy (Fig. 12.2). 

Allan, V. (2000). Protein localization by fluorescence microscopy A practical approach. Oxford University Press, Oxford, UK. 

Chen, Y., Mills, J. D. and Periasamy, A. (2003). Protein localization in living cells and tissues using FRET and FLIM. Differentiation 71, 528-41. 

However, engineering of fluorescent marker proteins to determine subcellular protein localizations and associations in planta can be quite challenging since plant cells contain a number of autofluorescent compounds (e.g., lignin, chlorophyll, phenols, etc.,) whose emission spectra interfere with that of the most commonly used green or red fluorophores and their spectral variants... 

It is important that the method used to detach cells from their growing surface is compatible with end use. For final use as cell control material, it is important to use a methodology that preserves structural integrity and membrane protein localization. Enzymatic-based reagents may affect proteins on the surface of cells. 

The last sample preparation method for IMS is the transfer of a tissue section onto the PVDF membrane. Proteins in the section can be transferred onto the PVDF membrane and then analyzed on the membrane. The advantage of this method is that the enzyme can be digested for MS" measurement, because the information on protein localization in the organization is fixed on the membrane.5,20 This technique can denature, reduce, and digest the proteins in the tissue section efficiently and remove the salt from the tissue. This increases the efficiency with which biological molecules are ionized, making it possible to obtain sensitive mass imaging spectra. 

The MBPs are extrinsic proteins localized exclusively at the cytoplasmic surface in the major dense line (Fig. 4-11), a conclusion based on their amino acid sequence, inaccessibility to surface probes and direct localization at the electron microscope level by immunocytochemistry. There is evidence to suggest that MBP forms dimers, and it is believed to be the principal protein stabilizing the major dense line of CNS myelin, possibly by interacting with negatively charged lipids. A severe hypomyelination and failure of compaction of the major dense line in MBP deficient shiverer mutants supports this hypothesis (Table 4-2). 

Allen, P. B., Ouimet, C. C. and Greengard, P. Spinophilin, a novel protein phosphatase 1 binding protein localized to dendritic spines. Proc. Natl Acad. Sci. U.S.A. 94 9956-9961, 1997. 

The Ras proteins are synthesized as biologically inactive, cytosolic precursor proteins. They are then modified by several post-translational processing steps at the carboxyl terminal end and thereby converted into biologically active proteins localized at the plasma membrane. The cysteine of the C-terminal CAAX sequence (C is cysteine, A is generally an aliphatic amino acid, and X is methionine, serine, alanine, or glutamine) is first enzymatically S-farnesylated the AAX part is then cleaved off by a specific protease, and the free C-terminal cysteine is finally converted into a methyl ester (Scheme 1). 

Synaptotagmins are yet another family of Ca2+-binding proteins, localized on the membranes of synaptic vesicles, where they seem to be involved in the release of neurotransmitters. While the mechanism by which they are involved in Ca2+-mediated synaptic transmission is unclear, it seems likely that the neurotoxicity of heavy metals such as Pb is due to a higher affinity of synaptotagmins for Pb2+ than for Ca2+. 

Sekar RB, Periasamy A (2003) Fluorescence resonance energy transfer (FRET) microscopy imaging of live cell protein localizations. J Cell Biol 160 629-33... 

For a complete functional study of a biological pathway, it is often necessary to confirm the important protein interactions by in vivo experiments. This can be done by demonstrating protein localizations on a microscopic level, for instance, by tagging proteins with the green fluorescent protein or localizing them with antibodies and colloidal gold particles using an electron microscope. Additional, very specific biochemical experiments are often required to confirm the putative protein function. 

If the /3-rich conformation of outer membrane proteins is really the determinant of their localization, the prediction system of protein localization should evaluate the possibility of an input protein being the [3 type. Fortunately, this appears easier than ordinary secondary structure prediction of globular proteins. Several authors have proposed prediction methods. Here, a method that is conceptually simple and two other recently published methods are briefly described. 

Burns, N., Grimwade, B., Ross-Macdonald, P., Choi, E.-Y., Finberg, K., Roeder, G., and Snyder, M. (1994). Large-scale analysis of gene expression, protein localization, and gene disruption in Saccharomyces cerevisiae. Genes Dev. 8, 1087—1105. 

Garcia-Bustos, J., Heitman, J., and Hall, M. (1991). Nuclear protein localization. Biochim. Biophys. Acta 1071, 83-101. 

Nakai, K., and Kanehisa, M. (1991). PROTEINS Structure, function, and genetics expert system for predicting protein localization sites in gram-negative bacteria. Proteins Struct. Fund. Genet. 11, 95-110. 

Shapiro, L. (1993). Protein localization and asymmetry in the bacterial cell. Cell 73, 841-855. 

Sjostrom, M., Wold, S., Wieslander, A., and Rilfors, L. (1987). Signal peptide amio acid sequences in Eschericha coli contain information related to final protein localization. A multivariate data analysis. EMBO J. 6, 823—831. 

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