tailieunhanh - Báo cáo khoa học: Mouse cytosolic sulfotransferase SULT2B1b interacts with cytoskeletal proteins via a proline⁄serine-rich C-terminus

Cytosolic sulfotransferase (SULT) SULT2B1b had previously been charac-terized as a cholesterol sulfotransferase. Like human SULT2B1, mouse SULT2B1b contains a unique, 31 amino acid C-terminal sequence with a proline⁄serine-rich region, which is not found in members of other SULT families. To gain insight into the functional relevance of this proline⁄ser-ine-rich region, we constructed a truncated mouse SULT2B1b lacking the 31 C-terminal amino acids, and compared it with the wild-type enzyme. | Mouse cytosolic sulfotransferase SULT2B1b interacts with cytoskeletal proteins via a proline serine-rich C-terminus Katsuhisa Kurogi1 Yoichi Sakakibara1 Yosuke Kamemoto1 Saki Takahashi1 Shin Yasuda2 Ming-Cheh Liu3 and Masahito Suiko1 1 Department of Biochemistry and Applied Biosciences University of Miyazaki Japan 2 Department of Bioscience Schoolof Agriculture Tokai University Aso Kumamoto Japan 3 Department of Pharmacology College of Pharmacy The University of Toledo OH USA Keywords cholesterol cytoskeleton interaction sulfation sulfotransferase Correspondence Y. Sakakibara Department of Biochemistry and Applied Biosciences University of Miyazaki 1-1 Gakuenkibanadai-Nishi Miyazaki Miyazaki 889-2192 Japan Fax Tel 81 985 58 7211 E-mail ysakaki@ Received 1 May 2010 revised 8 July 2010 accepted 15 July 2010 doi Cytosolic sulfotransferase SULT SULT2B1b had previously been characterized as a cholesterol sulfotransferase. Like human SULT2B1 mouse SULT2B1b contains a unique 31 amino acid C-terminal sequence with a proline serine-rich region which is not found in members of other SULT families. To gain insight into the functional relevance of this proline ser-ine-rich region we constructed a truncated mouse SULT2B1b lacking the 31 C-terminal amino acids and compared it with the wild-type enzyme. Enzymatic characterization indicated that the catalytic activity was not significantly affected by the absence of those C-terminal residues. Glutathione S-transferase pulldown assays showed that several proteins interacted with mouse SULT2B1b specifically through this C-terminal proline serine-rich region. Peptide mass fingerprinting revealed that of the five SULT2B1b-binding proteins analyzed three were cytoskeletal proteins and two were cytoskeleton-binding molecular chaperones. Furthermore wild-type mouse SULT2B1b but not the truncated enzyme was associated with the cytoskeleton in experiments with a cytoskeleton-stabilizing .