Abstract
Using two-dimensional (2-D) gel electrophoresis, human perilymph and cerebrospinal fluid have been shown to be highly enriched for an acidic protein with MR 30,000, we designated it as AP30. The protein exhibits charge heterogeneity, with at least eight isoforms visible between pI 4.5 to 5.5 on 2-D gels. Purification of the protein was carried out by ammonium sulfate precipitation, polybuffer exchanger column chromatofocusing, and acetone fractional precipitation. The resulting preparation also contains eight spots in the acidic area of 2-D gels, and one broad band located at Mr 30,000 by SDS-PAGE. Digestion of AP30 with neuraminidase causes the isoforms to shift to a more basic position and to consolidate into two primary spots, indicating that AP30 is a variably sialylated glycoprotein. Amino acid analysis of AP30 revealed an amino acid content very similar to that of human apolipoprotein D. Attempts to determine the amino acid sequence demonstrated that the N-terminus is blocked. Edman sequencing of two peptide fragments, generated by cyanogen bromide cleavage of AP30, both revealed sequences having 100% identity to human apolipoprotein D. Western blot analysis of AP30 with the antibody against authentic human apolipoprotein...Continue Reading
References
Feb 10, 1976·Biochemistry·W J McConathy, P Alaupovic
May 7, 1976·Analytical Biochemistry·M M Bradford
Apr 1, 1992·Electrophoresis·L Holmquist
Apr 1, 1991·Otolaryngology--head and Neck Surgery : Official Journal of American Academy of Otolaryngology-Head and Neck Surgery·D R PaughM J Disher
Jan 1, 1991·Progress in Lipid Research·P K WeechE Rassart
Nov 1, 1990·The Biochemical Journal·M BalbínC López-Otín
Jan 15, 1990·Biochemical and Biophysical Research Communications·W JahnenR J Simpson
Jan 1, 1990·European Archives of Oto-rhino-laryngology : Official Journal of the European Federation of Oto-Rhino-Laryngological Societies (EUFOS) : Affiliated with the German Society for Oto-Rhino-Laryngology - Head and Neck Surgery·E ArrerH J Gibitz
Jan 1, 1989·Experimental Cell Research·Q Sun
Jul 1, 1988·Acta Oto-laryngologica·E ArrerH J Gibitz
Jan 1, 1986·Methods in Enzymology·W J McConathy, P Alaupovic
Jan 1, 1973·Methods in Enzymology·H D Niall
Aug 15, 1970·Nature·U K Laemmli
Oct 1, 1966·Archives of Otolaryngology·H Silverstein, H F Schuknecht
Citations
Nov 5, 2003·Neuroscience·F Glöckner, T G Ohm
Feb 27, 2007·Hearing Research·Donald E ColingRichard J Salvi
Sep 9, 2004·Otology & Neurotology : Official Publication of the American Otological Society, American Neurotology Society [and] European Academy of Otology and Neurotology·John F McGuire, Begona Casado
Jun 27, 2001·Electrophoresis·P R BurkhardE Shiffer
Feb 19, 2000·Biochemical and Biophysical Research Communications·V N Trieu, F M Uckun
Oct 26, 2021·Frontiers in Physiology·Diego Sanchez, Maria D Ganfornina