. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 128 L. M. RZEPECKI ET 20 40 60 Elulion time (mini Figure 2. HPLC chromatography profiles of Mefp-2 (- and Mefp-l ( 1 monitored at 280 nm. Proteins were resolved on a x 25 cm semi-preparative C8 column with an acetonitrile gradient, as indicated by the inclined dashed line. Inset: AU-PAGE of punned Mefp- 2 at three concentrations, stained with Coomassie Blue. The arrow in- dicates the position of trace Mefp-l contaminants. mobility were also observed in this distal segment, but their origin and nature were not invest
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 128 L. M. RZEPECKI ET 20 40 60 Elulion time (mini Figure 2. HPLC chromatography profiles of Mefp-2 (- and Mefp-l ( 1 monitored at 280 nm. Proteins were resolved on a x 25 cm semi-preparative C8 column with an acetonitrile gradient, as indicated by the inclined dashed line. Inset: AU-PAGE of punned Mefp- 2 at three concentrations, stained with Coomassie Blue. The arrow in- dicates the position of trace Mefp-l contaminants. mobility were also observed in this distal segment, but their origin and nature were not investigated further. To obtain a better separation between phenol and accessory glands, we micro-dissected phenol and accessory glands from thin foot sections and again, Mefp-2 was exclusively detected in the phenol gland, whereas Mefp-l appeared in both glands. No other byssal glands are known to con- tain DOPA proteins. Extraction of byssal plaques in either acetic acid-urea or neutral SDS buffers yielded a polypeptide that co-mi- grated with Mefp-2 during both SDS- and AU-PAGE and stained positively with NET (Fig. Ib. c): miscellaneous electrophoretic species that stained with Coomassie Blue, but not NBT, also appeared. Preliminary amino acid analysis following HPLC purification of this NBT-positive plaque polypeptide revealed a composition very similar to authentic Mefp-2 (Diamond and Waite. unpub.). No polypeptide extracted from byssal threads alone exhibited NBT reactivity or electrophoretic behavior akin to Mefp- 2, and no polypeptides corresponding to intact Mefp-l were detected in plaque or thread extracts. Dot blots con- firmed that a considerable proportion (ca. 10%) of acid- soluble plaque protein was NBT-sensitive. and immu- noreactivity assays demonstrated that the extracted ma- terial that reacted with anti-Mefp-2 antibodies was specific to plaques (Fig. Id). Pre-immune serum was unreactive in these assays. These observations are complicated by some cross-reactivity b
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
