Clupea pallasii

Figure 10.4 Comparison of origins of C16-C22 monoethylenic fatty acids in a marine Canadian Pacific herring (Clupea pallasii) and a freshwater alewife (Canadian Great Lakes Alosa pseudoharengus). Adapted from Ackman (1982).

Japanese biochemists fractionated clupeine prepared from Clupea pallasii into two fractions, Y and Z, by elution chromatography on a column of alumina which had been buffered with the eluting solvent (0.45 M or 0.48 M K2HPO4, pH 9.0—9.2) (Ando and Sawada, 1959, 1961). On subsequent application of countercurrent distribution (solvent system -butanol vs, 0.12 M sodium -toluenesulfonate containing 0.40 M NaCl), the Y fraction was further separated into YI and YII, while the Z fraction was found to be homogeneous (see Chap. VII. B). 

In an attempt to trace the origin of the heterogeneity, clupeine and iridine specimens were prepared under exactly the same mild conditions from completely mature testes of individual specimens of Clupea pallasii (Ando and Sawada, 1959, 1962) and Salmo irideus (Ando and Sawada, 1959, 1960), respectively. They used three Pacific herring caught at three different places off Hokkaido and three rainbow trout obtained... 

Clupeine from Pacific herring (Clupea pallasii), Amino cid contents were estimated by densitometry of multiply developed paper chromatograms. ( + + + ) indicates the presence in a large amount and ( + ) in an appreciable amount, though not estimated. + + and + in appreciable amounts when estimated, only a trace and — absence. 

Four different batches of clupeine specimens from Clupea pallasii were separated by means of this chromatographic system. The fractions of the peaks of YI, YII and Z obtained were 34 6%, 36 5% and 26 4%, respectively. The amount of F, a minor fraction that was eluted before the YII fraction (cf. Figs. VII-7, 8), was 4 3%. Thus about 96% of whole clupeine is composed of the three main components, YI, YII and Z, and their average ratio in whole clupeine is 1 1 0.7s [Suzuki and Ando, 1968 (2)]. 

Whole clupeine from Clupea harengus was also fractionated by chromatography into three components, clupeine YT, YTI and Z (see Chap. VIII. E) (Chang, Nakahara and Ando, to be published), just like clupeine from Clupea pallasii. 

Fig. VII-8. Column chromatographic fractionation on preparative scale of protamines into all or some of their components using CM-Sephadex C-25 or Bio-Gel CM-2. A. Protamine ca. 25 mg of whole clupeine sulfate (from Clupea pallasii) in a small amount of water. Column 0.9 X 130 cm of Bio-Gel CM-2 (high capacity). Elution 0.05 M acetate buffer, pH 5.8, containing 1.5 M NaCl at room temperature at a flow rate of 2.4 ml/h. B. Protamine ca. 50 mg of whole salmine sulfate (from Oncorhynchus ketd) in a small amount of water. Column 0.9 X 150 cm of Bio-Gel CM-2 (high cap.). Elution 0.05 M acetate buffer, pH 5.8, containing 1.0 M and 1.5 M NaCl at room temperature at a flow rate of 10 ml/h. C Protamine ca. 600 mg of whole iridine sulfate (from Salmo irideus) in as small as possible amount of the eluting buffer containing 0.5 M NaCl. Column 3.0 x 146 cm of CM-Sephadex C-25. Elution 0.05 M acetate buffer, pH 5.8, containing 1.5 M NaCl at room temperature at a flow rate of 85 ml/b (Reproduced from Ando, T., and Watanabe, S., 1969)...

The primary and secondary structures of all the molecular species of clupeine (from Pacific herring, Clupea pallasii) YI, YII and Z, have been elucidated [see Chart VIII-6 for the primary structures, and Suzuki and Ando, 1968 (1), for the secondary structures]. This is the first and only instance at present of structure determination and elucidation of the number of molecular species present in the... 

Neither the exhaustive studies of the Tokyo group (1964 and 1969) as described in the next section (Chap. VIII. E) nor the partial study by Felix and Hashimoto (1963) have revealed any differences in the amino-acid sequences of the corresponding components of clupeine from Norwegian Sea and North Sea (Clupea harengus) and Pacific (Clupea pallasii) herring. 

However, all these results were obtained by analysis of unfractionated clupeine. The complete amino-acid sequences recently determined for the three components of clupeine from Pacific herring (Clupea pallasii) (Ando et al,y 1962 Ando and Suzuki, 1966, 1967) confirmed the presence of an amino-acid residue, glycine, as a constituent and of the N-terminal alanine in addition to the N-terminal proline in some of the clupeine molecules. At that time no complete fractionation of clupeine from Clupea harengus into a number of homogeneous components had been accomplished. 

Ando, T., Abukumagawa, E., Nagai, Y., Yamasaki, M. (2) On the N-terminal residues and sequence of clupeine from Clupea pallasii The occurrence of proline in addition to alanine as the N-terminus. J. Biochem. (Tokyo) 44, 191—194 (1957). 

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