. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 140 L. H. KLEINHOLZ ET AL. C 14r min E c s <M Q O 20 r 10. X O Q. 10 15 20 Time ( Minutes) 25 30 FIGURE 4. The major active zone from partition chromatography (fractions 55-60, Fig. 3) was subjected to reversed-phase HPLC on a column of Partisil-lO ODS-2 ( mm X 25 cm, Whatman) utilizing Waters Liquid Chromatography System (Data Module 730; Programmable Controller 721; Pumps 510; Absorbance Detector 441). Solvent system: linear gradient elution for 30 min from 30% CH3CN to 45% CH3CN, both wit
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 140 L. H. KLEINHOLZ ET AL. C 14r min E c s <M Q O 20 r 10. X O Q. 10 15 20 Time ( Minutes) 25 30 FIGURE 4. The major active zone from partition chromatography (fractions 55-60, Fig. 3) was subjected to reversed-phase HPLC on a column of Partisil-lO ODS-2 ( mm X 25 cm, Whatman) utilizing Waters Liquid Chromatography System (Data Module 730; Programmable Controller 721; Pumps 510; Absorbance Detector 441). Solvent system: linear gradient elution for 30 min from 30% CH3CN to 45% CH3CN, both with trifluoroacetic acid; flow rate, 1 ml/min. Bioassays were done with of each fraction; each aliquot was lyophilized and dissolved in ml saline; injected 50 ^I/crab into 5 test Uca. The reference compounds utilized were: synthetic PDH of Pandalus borealis (Riehm and Rao, 1982), retention time min (broken arrow); and synthetic PDH of Uca (Rao et al., 1985), retention time min (solid arrow). The PDH activity is expressed as Standard Integrated Response values, as denned by Fingerman et al. (1967). Studies involving paper electrophoresis (Stephens et al., 1956; Fingerman, 1966), disc electrophoresis (Rao et al., 1970; Keller, 1977), ion-exchange chromatography (Raoetal., 1970; Fernlund, 1971; Kleinholz, 1970, 1972; Dores and Herman, 1980), and reversed-phase HPLC (Keller and Kegel, 1984) showed that multiple forms of PDH are present in extracts of eyestalks or sinus glands of diverse decapod Crustacea. Initial resolution of the multiple forms of PDH can be best achieved, however, by ion- exchange chromatography (Kleinholz, 1970; Rao et al., 1985). Since the PDHs sep- arable by ion-exchange chromatography display identical elution profiles in gel filtra- tion, the multiple forms of PDH are thought to differ in net charge but not in molecular size (Kleinholz, 1972). This view is supported by the finding that the two sequenced PDHs are both octadecapeptides t
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