Antibodies from a 2-h chase onwards. An equivalent His-tagged, i.e.
Antibodies from a 2-h chase onwards. An equivalent His-tagged, i.e. C-terminal, ARSK-derived 23-kDa fragment could possibly be detected in Western blot analyses of ARSK enriched from conditioned medium of producer cells. Corresponding N-terminal fragment(s) could not be detected. They could possibly have escaped our analyses around the basis of antibody recognition due to incompatible epitopes following processing. Additional research on this challenge will demand expression of larger amounts of ARSK and/or availability of other ARSKspecific antibodies. ARSK is expressed in all tissues examined in this research and was also identified in eight tissues from rat in M6P glycoproteome analyses (33). Its ubiquitous expression pattern may perhaps suggest a typical and widespread sulfated substrate and signifies that ARSK deficiency probably leads to a lysosomal storage disorder, as shown for all other lysosomal sulfatases. Presently, we are generating an ARSK-deficient mouse model that really should pave the way to recognize the physiological substrate of this sulfatase and its general pathophysiological relevance. Finally, the mouse model could enable us to draw conclusions on ARSKdeficient human sufferers who so far escaped diagnosis and may well be available for enzyme substitute therapy. The presence of M6P on ARSK qualifies this sulfatase for such a treatment, which has established helpful for therapy of various other lysosomal storage problems.Acknowledgments–We thank Bernhard α2β1 manufacturer Schmidt and Olaf Bernhard for mass spectrometry; Nicole Tasch, Annegret Schneemann, Britta Dreier, Martina Balleininger (all from G tingen), William C. Lamanna, Jaqueline Alonso Lunar, Kerstin B er, and Claudia Prange for technical help; Markus Damme for preliminary evaluation of subcellular localization; and Jeffrey Esko (San Diego) for critically reading the manuscript. We also thank Kurt von Figura for help throughout the preliminary phase of this venture.Dierks, T. (2007) The heparanome. The enigma of encoding and decoding heparan sulfate sulfation. J. Biotechnol. 129, 290 07 Schmidt, B., Selmer, T., Ingendoh, A., and von Figura, K. (1995) A novel amino acid modification in sulfatases that is definitely defective in several sulfatase deficiency. Cell 82, 27178 von B ow, R., Schmidt, B., Dierks, T., von Figura, K., and Us , I. (2001) Crystal framework of an enzyme-substrate complex supplies insight in to the interaction amongst human arylsulfatase A and its substrates through catalysis. J. Mol. Biol. 305, 269 77 Dierks, T., Lecca, M. R., Schlotterhose, P., Schmidt, B., and von Figura, K. (1999) Sequence determinants directing conversion of cysteine to formylglycine in eukaryotic sulfatases. EMBO J. 18, 2084 091 Dierks, T., Schmidt, B., and von Figura, K. (1997) Conversion of cysteine to formylglycine. A protein modification in the endoplasmic reticulum. Proc. Natl. Acad. Sci. U.S.A. 94, PI3KC2β MedChemExpress 119631968 Dierks, T., Dickmanns, A., Preusser-Kunze, A., Schmidt, B., Mariappan, M., von Figura, K., Ficner, R., and Rudolph, M. G. (2005) Molecular basis for a number of sulfatase deficiency and mechanism for formylglycine generation with the human formylglycine-generating enzyme. Cell 121, 54152 Dierks, T., Schmidt, B., Borissenko, L. V., Peng, J., Preusser, A., Mariappan, M., and von Figura, K. (2003) Many sulfatase deficiency is triggered by mutations within the gene encoding the human C( )-formylglycine generating enzyme. Cell 113, 435444 Dierks, T., Schlotawa, L., Frese, M. A., Radhakrishnan, K., von Figura, K., and Schmidt, B. (2009) Molecular basi.
