Subcellular Location and Molecular Mobility of Human Cytosolic Sulfotransferase 1C1 in Living Human Embryonic Kidney 293 Cells

Cytosolic sulfotransferases were first isolated from the hepatic cytosol, and they have been localized in the cytoplasm of formaldehyde-fixed human cell samples. The current work was carried out to determine the subcellular localization and molecular mobility of cytosolic sulfotransferases in living human embryonic kidney (HEK) 293 cells. In this work, the subcellular location of human cytosolic sulfotransferase 1C1 (SULT1C1) was studied in cultured HEK293 cells using confocal laser-scanning microscopy. A green fluorescent protein (GFP)-tagged SULT1C1 protein was localized in the cytoplasm of living HEK293 cells. This is consistent with results from previous studies on several other cytosolic sulfotransferase isoforms. Fluorescence recovery after photobleaching microscopy was performed to assess the molecular mobility of the expressed GFP-SULT1C1 molecules. The results suggested that the expressed recombinant GFP-SULT1C1 molecules in living HEK293 cells may include both mobile and immobile populations. To obtain additional insights into the subcellular location of SULT1C1, two machine learning algorithms, Sequential Minimal Optimization and Multilayer Perceptron, were used to compute the probability distribution for the localization of SULT1C1 in nine selected cellular compartments. The resulting probability distribution suggested that the most likely subcellular location of SULT1C1 is the cytosol.

[1]  Hong Gu,et al.  A novel method for predicting protein subcellular localization based on pseudo amino acid composition. , 2010, BMB reports.

[2]  Nilanjan Roy,et al.  SubCellProt: Predicting Protein Subcellular Localization Using Machine Learning Approaches , 2009, Silico Biol..

[3]  Chittibabu Guda,et al.  TARGET: a new method for predicting protein subcellular localization in eukaryotes , 2005, Bioinform..

[4]  Richard N. Day,et al.  Imaging protein behavior inside the living cell , 2005, Molecular and Cellular Endocrinology.

[5]  C. Falany.  Enzymology of human cytosolic sulfotransferases , 1997, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[6]  R. Weinshilboum,et al.  Sulfotransferase molecular biology: cDNAs and genes , 1997, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[7]  P. Zhao,et al.  Combining Machine Learning and Homology-Based Approaches to Accurately Predict Subcellular Localization in Arabidopsis1[C][W][OA] , 2010, Plant Physiology.

[8]  S. Subramaniam,et al.  pTARGET [corrected] a new method for predicting protein subcellular localization in eukaryotes. , 2005, Bioinformatics.

[9]  J. Tischfield,et al.  Single-base deletion induced by benzo[a]pyrene diol epoxide at the adenine phosphoribosyltransferase locus in human fibrosarcoma cell lines. , 1994, Mutation research.

[10]  M. Whitnall,et al.  Estrogen and hydroxysteroid sulfotransferases in guinea pig adrenal cortex: cellular and subcellular distributions. , 1993, Endocrinology.

[11]  K. Yoshinari,et al.  Molecular characterization of ST1C1-related human sulfotransferase. , 1998, Carcinogenesis.

[12]  D. Ziegler,et al.  The enzymes of detoxication. , 1983, The Journal of biological chemistry.

[13]  D. Nimmagadda,et al.  Cellular Localization Studies on Human Estrogen Sulfotransferase SULT1E1 in Human Embryonic Kidney 293 Cells , 2007, Drug Metabolism and Disposition.

[14]  R. Neft,et al.  Genetic toxicity of 2-acetylaminofluorene, 2-aminofluorene and some of their metabolites and model metabolites. , 1994, Mutation research.

[15]  R. J. Anderson,et al.  Sulfation of iodothyronines by human sulfotransferase 1C1 (SULT1C1)*. , 2000, Biochemical pharmacology.

[16]  Vladimir Vapnik,et al.  The Support Vector Method , 1997, ICANN.

[17]  Peter J. Schaap,et al.  Molecular characterization of the , 1997 .

[18]  C. Falany,et al.  Expression and Localization of Cytosolic Sulfotransferase (SULT) 1A1 and SULT1A3 in Normal Human Brain , 2009, Drug Metabolism and Disposition.

[19]  J. Gardy,et al.  Methods for predicting bacterial protein subcellular localization , 2006, Nature Reviews Microbiology.

[20]  Rolf Apweiler,et al.  The SWISS-PROT protein sequence database and its supplement TrEMBL in 2000 , 2000, Nucleic Acids Res..

[21]  K. Nagata,et al.  Pharmacogenetics of sulfotransferase. , 2000, Annual review of pharmacology and toxicology.

[22]  Peer Bork,et al.  Predicting protein cellular localization using a domain projection method. , 2002, Genome research.

[23]  L. Pedersen,et al.  Structure and function of sulfotransferases. , 2001, Archives of biochemistry and biophysics.

[24]  J. Platt.  Sequential Minimal Optimization : A Fast Algorithm for Training Support Vector Machines , 1998 .

[25]  R. Weinshilboum,et al.  Aggresome formation and pharmacogenetics: sulfotransferase 1A3 as a model system. , 2004, Biochemical and biophysical research communications.

[26]  M. W Gardner,et al.  Artificial neural networks (the multilayer perceptron)—a review of applications in the atmospheric sciences , 1998 .

[27]  Ian H. Witten,et al.  Data mining: practical machine learning tools and techniques with Java implementations , 2002, SGMD.

[28]  Chittibabu Guda,et al.  Predicting the Subcellular Localization of Human Proteins Using Machine Learning and Exploratory Data Analysis , 2006, Genom. Proteom. Bioinform..

[29]  Mamoon Rashid,et al.  Support Vector Machine-based method for predicting subcellular localization of mycobacterial proteins using evolutionary information and motifs , 2007, BMC Bioinformatics.