. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. SENSORY TRANSDUCTION IN LARVAL CILIA 149. Figure 1. Scanning electron micrographs of the cilia purified from Haliotis larvae. (A) Scanning electron micrograph of cilia preparation (2400X). Contour lengths of the long propulsive cilia > ^m. (B) Isolated view of smaller proposed sensory cilia (length ca. ,um) in the same preparation (4500 • ). Rodbard and Munson (McPherson, 1985). Competition for binding by unlabelled L-lysine, L-ornithine, or L-a,fS- diaminopropionic acid was analyzed similarly, with the same progr
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. SENSORY TRANSDUCTION IN LARVAL CILIA 149. Figure 1. Scanning electron micrographs of the cilia purified from Haliotis larvae. (A) Scanning electron micrograph of cilia preparation (2400X). Contour lengths of the long propulsive cilia > ^m. (B) Isolated view of smaller proposed sensory cilia (length ca. ,um) in the same preparation (4500 • ). Rodbard and Munson (McPherson, 1985). Competition for binding by unlabelled L-lysine, L-ornithine, or L-a,fS- diaminopropionic acid was analyzed similarly, with the same program. ADP-ribosylation Cholera toxin-catalyzed ADP-ribosylation for the la- beling of G protein was performed as described by Pace and Lancet (1986), using cholera toxin preactivated by incubation with 20 mM dithiothreitol at room tempera- ture for 15 min. Cilia (200 jug/ml protein) were incubated at room temperature in a buffer containing 20 mM Tris- HC1 (pH ), 30 mM thymidine, 1 mM ATP, mA/ GTP, 5 mA/MgCl:, 1 mM EDTA, 3 mA/ phosphoenol- pyruvate, 5 units/ml pyruvate kinase, Triton X- 100, 1 mA/dithiothreitol, 5 //A/ [32P]-NAD (10-20 Ci/ mmol), and preactivated cholera toxin at a final concen- tration of 10 /ig/ml in a final volume of 200 n\. The re- action was terminated after 30 min by adding an equal volume of 2X electrophoresis sample buffer (100 mA/Tris- HC1 pH , 20% glycerol, 2% SDS, 100 mA/dithiothre- itol, and bromophenol blue) and boiling for 20 min. ADP ribosylation of cilia for G protein activation was conducted under similar conditions without radio- isotope (Pace and Lancet, 1986), using a 1-h incubation. Cilia then were centrifuged at 15,000 X # for 10 min at 4°C, resuspended in buffer, and assayed as described. Gel electrophoresis and autoradiography Proteins were separated by one-dimensional SDS poly- acrylamide gel electrophoresis (SDS-PAGE) (Laemmli. 1970) with a 10% separating gel. Molecular weight stan- dards were included on each ge
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