Conformational energy and circular dichroism computed for cyclo-(pro-gly)3

An analytic cyclization procedure has allowed the complete specification of c-(Pro-Gly)₃ conformation by three dihedral angles. Utilizing this simplification, the intramolecular potential energy of c-(Pro-Gly)₃ has been computed over all of conformational space. Conformers with all peptide bonds trans have the lowest total potential energy. Circular dichroism spectra calculated for the low-energy regions provide the basis for the interpretation of experimental CD spectra.     

Promotion of crown-gall tumor initiation on primary pinto bean leaves by certain inorganic salts

Crown-gall tumor initiation by Agrobacterium tumefaciens (SMITHand TOWN.) CONN, strain B6, on Phaseolus vulgaris L. var. "Pinto"was found to be sensitive to the addition of various salts tothe freshly inoculated primary leaves. Calcium, magnesium, zincand ammonium sulfates, cobalt and nickel chlorides and potassiumnitrate increased the number of tumors on inoculated leavesby 100 to 200 percent. Maximal promotions were obtained at concentrationsof 10^–7 to 10^–5 M with all of these salts exceptpotassium nitrate which was most active at 10^–2M. Sodium,potassium, calcium, magnesium and aluminum chlorides, and sodium,potassium and nickel sulfates had little or no effect on tumorinitiation. The combination of sodium sulfate with magnesiumchloride gave promotions comparable to those obtained with magnesiumsulfate, indicating that both the magnesium and sulfate ionswere necessary for the promotion obtained with this latter salt.Combining any two of the active salts at their optimum concentrationsfor promotion resulted in a reciprocal inhibition of the promotionobtained with either salt alone, suggesting these salts to beactive by different and potentially antagonistic mechanisms.The possible nature of these promotions is discussed, but theyremain too obscure to warrant a specific proposal for theirmode of action. ¹Present adress: Department of Botany, University of Khartoum,Khartoum, Sudan.     

Ultrastructural Study of the Development of Pleistophora schubergi Zwölfer, 1927 (Protozoa,Microsporida) in Larvae of the Spruce Budworm,Choristoneura fumiferana and Its Subsequent Taxonomic Change to the Genus Endoreticulatus

This study demonstrates that Pleistophora schubergi Zwölfer, 1927, a microsporidium originally isolated from the midgut epithelium of Nygmia phaeorrhoea Don (Euproctis chrysorrhoea L.) and Porthetria dispar L., and subsequently reported in several other insects including the spruce budworm, Choristoneura fumiferana (the host used in this investigation), does not belong in the genus Pleistophora Gurley, 1893. Pleistophora schubergi lacks the major features that are characteristic of Pleistophora typicalis, the type species of this genus. A comparison of ultrastructural observations reported for the type species of the genus Pleistophora, P. typicalis, and our observations of P. schubergi revealed significant differences. A thick (0.5 μm) amorphous coat, derived from parasite secretions and deposited external to the parasite plasmalemma, surrounds all developmental stages in P. typicalis. Double membranes, derived from host rough endoplasmic reticulum cisternae encircle the parasite plasmalemma of all developmental stages in P. schubergi. The sporophorous vesicle encases the spores in P. typicalis, and originates from the parasite-secreted coat that is present around meronts. In P. schubergi, the host endoplasmic reticulum cisternae form the envelope that surrounds the meronts. Moreover, the sporophorous vesicle envelope in P. typicalis persists around groups of spores, while in P. schubergi this envelope breaks easily to release the spores in the host cytoplasm. By comparing the characteristics of the microsporidium found in the spruce budworm with those of the recently created polysporous genera that sporulate within a vesicle, we found that P. schubergi does belong in the new genus Endoreticulatus Brooks et al. 1988, and consequently rename it Endoreticulatus schubergi (Zwölfer, 1927) n. comb.     

Use of the heterobifunctional cross-linker m-maleimidobenzoyl N-hydroxysuccinimide ester to affinity label cholecystokinin binding proteins on rat pancreatic plasma membranes

The binding of 125I-cholecystokinin-33 (125I-CCK-33) to its receptors on rat pancreatic membranes was decreased by modification of membrane protein sulfhydryl groups. Sulfhydryl modifying reagents also caused an accelerated release of bound 125I-CCK-33 from its receptor. Because of the presence of an essential sulfhydryl group(s) in CCK receptor binding we studied the application of the heterobifunctional (SH,NH2) cross-linker, m-maleimidobenzoyl N-hydroxysuccinimide ester (MBS), to affinity label 125I-CCK-33 binding proteins on rat pancreatic plasma membranes. Analysis of the cross-linked products by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography revealed that this heterobifunctional cross-linker affinity labeled a major Mr = 80,000-95,000 protein previously identified as part of the CCK receptor on the basis of affinity labeling using homobifunctional and heterobifunctional photoreactive cross-linkers. Additional proteins of Mr greater than 200,000, and Mr = 130,000-140,000 were affinity labeled using MBS. The efficiency of the cross-linking reaction between 125I-CCK-33 and its membrane binding proteins with MBS was significantly greater than that obtained with NH2-directed homobifunctional reagents such as disuccinimidyl suberate. The efficiency of cross-linking could be dramatically improved by reduction of membrane proteins with low-molecular weight thiols prior to binding and cross-linking. The differential labeling patterns of the CCK binding proteins obtained with chemical cross-linkers of similar length but different chemical reactivity underscores the need for caution in predicting native receptor structure from affinity labeling data alone. Using the same pancreatic plasma membrane preparation and 125I-insulin, the Mr = 125,000 alpha-subunit of the insulin receptor was affinity labeled using MBS as cross-linker, demonstrating its utility in identifying other peptide hormone receptors.     

Effects of exogenous methionine on storage protein composition of soybean cotyledons cultured in vitro

Supplemental methionine in a complete culture medium increased the methionine content of the protein fraction of cultured soybean (Glycine max L. Merrill) cotyledons (Thompson, Madison, Muenster 1981 Phytochemistry 20: 941-945). To explain the observed increase in protein methionine, we have measured the amounts and subunit compositions of 7S and 11S storage proteins and determined the amino acid compositions of the three major protein fractions (2-5S, 7S, 11S) of seeds developed on plants and of cultured cotyledons grown in the presence or absence of supplemental l-methionine. Development of cultured cotyledons was representative of development of seeds on plants. The ratios of 11S to 7S proteins, the subunit contents, and amino acid compositions of their storage protein fractions were similar, but not identical. Supplemental methionine increased the mole percent methionine in each of the three protein fractions of cultured cotyledons and changed the amounts of several other amino acids. Supplemental methionine inhibited expression of the 7S beta-subunit gene. Concomitant with the absence of the beta-subunit, which contains no methionine, was an increase in the ratio of 11S to 7S proteins, and an increase in the methionine content of the subunits composing these fractions. Inhibition of beta-subunit gene expression by methionine in cultured cotyledons provides a reproducible, easily controlled system for the study of eucaryotic gene expression.     

On the Composition, Deposition and Mobilization of Proteins in the Cotyledons of Castor Bean (Ricinus communis L. cv. Hale) Seeds: Their Role as Storage Proteins

Proteins in the soluble and insoluble fractions, extracted frommature castor bean cv. Hale seed cotyledons, differ quantitativelyand qualitatively from their counterparts extracted from theendosperm. The soluble fraction contains no glycoproteins, andthe lectins RCA₁ and ricin D are absent. While the insolubleproteins are electrophoretically and immunologically similarto those in the endosperm, they do not form the 100 kD subunitdimers which characterize some of the endosperm insoluble crystalloidproteins. Rapid rates of deposition of all of the soluble andinsoluble proteins present in the mature seed cotyledons commences30–35 d after pollination (DAP) and continues until 45DAP. These proteins are mobilized rapidly beginning 1–2d after seed imbibition and this coincides with an increasein specific activity, in the cotyledons, of two aminopeptidasesand a carboxypeptidase. The soluble and insoluble proteins inthe cotyledons of the mature seed probably function as storageproteins and support the growth of the germinated seed priorto the mobilization of the major protein storage reserves ofthe endosperm. Key words: Ricinus communis, Castor bean, Hale cultivar, Cotyledon, Storage protein, Seed development, Seed germination     

Effect of parental mate choice and semi‐natural early rearing environment on the growth performance and seawater tolerance of Chinook salmon Oncorhynchus tshawytscha

To assess whether parental mate choice and early rearing in a semi‐natural spawning channel may benefit the culture of Chinook salmon Oncorhynchus tshawytscha, 90 day growth trials were conducted using hatchery O. tshawytscha (hatchery), mate choice O. tshawytscha (i.e. the offspring of parents allowed to choose their own mate) that spent 6 months in a spawning channel prior to hatchery rearing (channel) and mate choice O. tshawytscha transferred to the hatchery as fertilized eggs (transfer). During the growth trials, all O. tshawytscha stocks were reared separately or in either mixed channel and hatchery or transfer and hatchery groups for comparison of performance to traditional practices. After 60 days in fresh water, all O. tshawytscha were transferred to seawater for an additional 30 days. Reared separately, all stocks grew c. 4·5 fold over 90 days but specific growth rate (G) and food conversion efficiency were higher in fresh water than after seawater transfer on day 60. In contrast, hatchery O. tshawytscha from mixed hatchery and channel and hatchery and transfer growth trials had a larger mass and length gain than their counterparts on day 60, but reduced G in seawater. In general, plasma levels of growth hormone, insulin‐like growth factor I and cortisol did not differ among any O. tshawytscha groups in either the separate or mixed growth trials. Despite some differences in gill Na⁺,K⁺‐ATPase activity, all O. tshawytscha had a high degree of seawater tolerance and experienced virtually no perturbation in plasma chloride following seawater transfer. Overall, all O. tshawytscha exhibited similar growth and seawater performance under traditional hatchery conditions and any benefit derived from either parental mate choice or semi‐natural early rearing environment was only observed in the presence of mutual competition with hatchery O. tshawytscha.     

New views of humpback whale flow dynamics and oral morphology during prey engulfment

The rise of inexpensive, user‐friendly cameras and editing software promises to revolutionize data collection with minimal disturbance to marine mammals. Video sequences recorded by aerial drones and GoPro cameras provided close‐up views and unique perspectives of humpback whales engulfing juvenile salmon at or just below the water surface in Southeast Alaska and Prince William Sound. Although humpback feeding is famous for its flexibility, several stereotyped events were noted in the 47 lunges we analyzed. Engulfment was rapid (mean 2.07 s), and the entrance through which the tongue inverts into the ventral pouch was seen as water rushes in. Cranial elevation was a major contributor to gape, and pouch contraction sometimes began before full gape closure, with reverberating waves indicating rebounding flow of water within the expanded pouch. Expulsion of filtered water began with a small splash at the anterior of the mouth, followed by sustained excurrent flow in the mouth's central or posterior regions. Apart from a splash of rebounding water, water within the mouth was surprisingly turbulence‐free during engulfment, but submersion of the whale's head created visible surface whirlpools and vortices which may aggregate prey for subsequent engulfment.     

Reproductive isolation following reintroduction of Chinook salmon with alternative life histories

We evaluated reproductive isolation of Chinook salmon (Oncorhynchus tshawytscha) life history types that have been reintroduced to northern Idaho, USA. Analysis of 1003 samples at six microsatellite loci revealed strong reproductive isolation between ocean- and stream-type Chinook salmon (fall and summer spawn timing, respectively) within the Clearwater River sub-basin (F _ST = 0.148, P < 0.00001). Very little evidence for gene flow among the two life history types was observed as assignment tests correctly assigned 99.6% of individuals in reference collections to either ocean- and steam-type Chinook salmon. Assignment of naturally reared juveniles indicated that both life history types were present with 24.1% stream-type and 75.9% ocean-type. Previous studies suggest high levels of divergence among the two life history types in natural populations, and our study verifies the persistence of reproductive isolation among types following colonization of habitat. Successful colonization of new habitat by (re)introduced species is likely influenced by diversity in life history types and this strategy has lead to naturally spawning populations in a variety of available habitats in the Clearwater River. As many populations of O. tshawytscha are listed as threatened or endangered under the U.S. Endangered Species Act, hope for recovery lies not only in effective management and habitat improvement, but adaptability of this species.