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 may 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 may well have escaped our analyses around the basis of antibody recognition due to incompatible epitopes right after processing. Additional research on this problem will demand expression of larger amounts of ARSK and/or availability of other ARSKspecific antibodies. ARSK is expressed in all tissues examined within this review and was also identified in eight tissues from rat in M6P glycoproteome analyses (33). Its ubiquitous expression pattern may possibly recommend a widespread and widespread sulfated substrate and indicates that ARSK deficiency almost certainly results in a lysosomal storage disorder, as shown for all other lysosomal sulfatases. At present, we are creating an ARSK-deficient mouse model that must pave the solution to determine the physiological substrate of this sulfatase and its all round pathophysiological relevance. Ultimately, the mouse model could allow us to draw conclusions on ARSKdeficient human patients who up to now escaped diagnosis and could be available for enzyme substitute therapy. The presence of M6P on ARSK qualifies this sulfatase for such a therapy, which has established valuable for therapy of a lot of other lysosomal storage disorders.Acknowledgments–We thank Bernhard Schmidt and Olaf Bernhard for mass spectrometry; Nicole Tasch, Annegret Schneemann, Britta Dreier, Martina Balleininger (all from G PLK1 manufacturer tingen), William C. Lamanna, Jaqueline Alonso Lunar, Kerstin B er, and Claudia Prange for technical assistance; Markus Damme for initial evaluation of subcellular localization; and Jeffrey Esko (San Diego) for critically studying the manuscript. We also thank Kurt von Figura for assistance in the course of the first 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 certainly defective in a number of 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 gives insight in to the interaction among human arylsulfatase A and its substrates for the duration of 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 5-HT3 Receptor Agonist manufacturer protein modification within the endoplasmic reticulum. Proc. Natl. Acad. Sci. U.S.A. 94, 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 various sulfatase deficiency and mechanism for formylglycine generation on 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) Multiple sulfatase deficiency is caused by mutations inside the gene encoding the human C( )-formylglycine producing enzyme. Cell 113, 435444 Dierks, T., Schlotawa, L., Frese, M. A., Radhakrishnan, K., von Figura, K., and Schmidt, B. (2009) Molecular basi.
