Aperiplasmic protein (Skp) of Escherichia coli selectively binds a class of outer membrane proteins

A search was performed for a periplasmic molecular chaperone which may assist outer membrane proteins of Escherichia coli on their way from the cytoplasmic to the outer membrane. Proteins of the periplasmic space were fractionated on an affinity column with sepharose-bound outer membrane porin OmpF. A 17kDa polypeptide was the predominant protein retained by this column. The corresponding gene was found in a gene bank; it encodes the periplasmic protein Skp. The protein was isolated and it could be demonstrated that it bound outer membrane proteins, following SDS-PAGE, with high selectivity. Among these were OmpA, OmpC, OmpF and the maltoporin LamB. The chromosomal skp gene was inactivated by a deletion causing removal of most of the signal peptide plus 107 residues of the 141-residue mature protein. The mutant was viable but possessed much-reduced concentrations of outer membrane proteins. This defect was fully restored by a plasmid-borne skp gene which may serve as a periplasmic chaperone.     

Effects of nitrogen on Pinus palustris foliar respiratory responses to elevated atmospheric CO2 concentration

Indirect effects of atmospheric CO₂ concentration [CO₂], onlongleaf pine (Pinus palustris Mill.) foliage respiration werestudied by growing trees in a factorial arrangement of low andhigh [CO₂] (369 and 729µmol CO₂ mol^–1) and low andhigh N (40 and 400 kg ha^–1 yr^–1). Direct effectsof [CO₂] on leaf respiration were tested by measuring respirationrates of foliage from all treatments at two CO₂ levels (360and 720µmol CO₂mol^–1) at the time of measurement.Elevated CO₂ did not directly or indirectly affect leaf respirationwhen expressed on a leaf area or mass basis, but a significantincrease in respiration per unit leaf N was observed in treesgrown in elevated [CO₂] (indirect response to elevated [CO₂]).The lack of a [CO₂] effect on respiration, when analysed onan area or mass basis, may have resulted from combined effectsof [CO₂] on factors that increase respiration (e.g. greateravailability of non-structural carbohydrates stimulating growthand carbon export from leaves) and on factors that decreaserespiration (e.g. lower N concentration leading to lower constructioncosts and maintenance requirements). Thus, [CO₂] affected factorsthat influence respiration, but in opposing ways. Key words: Pinus palustris, elevated CO₂, nitrogen, foliar, respiration     

A new periplasmic protein of Escherichia coli which is synthesized in spheroplasts but not in intact cells.

The gene spy from Escherichia coli has been cloned and sequenced. It encodes a precursor of a so far unknown 139-residue, rather basic periplasmic protein. It was not detectable immunologically in intact cells but was produced abundantly in spheroplasts. It could be a stress protein specific for spheroplasting.     

The haustorium of the root parasite Triphysaria (Scrophulariaceae), with special reference to xylem bridge ultrastructure

Haustoria of Triphysaria pusilla and T. versicolor subsp. faucibarbata from a natural habitat were analyzed by light and electron microscopy. Secretory trichomes (root hairs) participate in securing the haustorium to the surface of the host root. The keel-shaped intrusive part of the secondary haustorium penetrates to the depth of the vascular tissue of the host. Some of the epidermal interface cells differentiate into xylem elements. A significant number of haustoria do not differentiate further, but in most haustoria one to five of the epidermal xylem elements terminate a similar number of xylem strands. The strands mostly consist of vessel members and they connect host xylem or occasionally host parenchyma to the plate xylem adjacent to the stele of the parasite root. Each strand of this xylem bridge is accompanied by highly protoplasmic parenchyma cells with supposed transfer cell function. Increased surface area of the plasmalemma occurs in these cells as it does in interface parenchyma cells. Graniferous tracheary elements are restricted to the haustorium and occur most frequently in the plate xylem. The plate xylem is also accompanied by highly protoplasmic parenchyma cells. Hyphae of mycorrhizal fungi of the host root occasionally penetrate into the distal part of the xylem bridge. We combine structural observations and physiological facts into a hypothesis for translocation of water and nutrients between host and parasite. Some evolutionary aspects related to endogeny/exogeny of haustoria are discussed, and it is argued that the Triphysaria haustorium represents a greatly advanced and/or reduced condition within Scrophulariaceae.     

Biotransformation of diphenyl ether by the yeastTrichosporon beigelii SBUG 752

Trichosporon beigelii SBUG 752 was able to transform diphenyl ether. By TLC, HPLC, GC, GC-MS, NMR- and UV-spectroscopy, several oxidation products were identified. The primary attack was initiated by a monooxygenation step, resulting in the formation of 4-hydroxydiphenyl ether, 2-hydroxydiphenyl ether and 3-hydroxydiphenyl ether (48:47:5). Further oxidation led to 3,4-dihydroxydiphenyl ether. As a characteristic product resulting from the cleavage of an aromatic ring, the lactone of 2-hydroxy-4-phenoxymuconic acid was identified. The possible mechanism of ring cleavage to yield this metabolite is discussed.     

Constrained C-terminal hexapeptide neurotensin analogues containing a 3-oxoindolizidine skeleton

Summary In order to enforce different spatial orientations in the C-terminal hexapeptide of neurotensin (NT_8–13) and to gain information about the importance of the 10–11 peptide bond for binding to NT receptors, the Pro¹⁰-Tyr¹¹ fragment has been replaced with (2R,8S,8aR)-, (2S,8S,8aR)-, (2S,8S,8aS)-, (2S,8R,8aS)- and (2R,8R,8aS)-8-amino-2-benzyl-3-oxoindolizidine-2-carboxylic acid. Molecular dynamics calculations and energy minimization studies have shown that, contrarily to the Pro-Tyr moiety, none of these indolizidines display a tendency to adopt type I and III -turns, but those having (8S,8aR) or (8R,8aS) stereochemistry essentially adopt extended conformations and the (8S,8aS) stereoisomer prefers a nonstandard folding. The four diastereomeric NT_8–13 analogues incorporating (8S,8aR) or (8R,8aS) indolizidines displayed binding affinities for the brain NT receptor similar to that of [Ala¹¹]-NT_8–13 and only five- to ninefold lower than that of the corresponding analogue, [Phe¹¹]NT_8–13. Although this slight decrease could be attributed to differences in conformational behavior between these constrained NT_8–13 analogues and [Phe¹¹]NT_8–13 or NT_8–13, it is not clear whether the -turn around Pro¹⁰-AA¹¹ (AA=Phe, Tyr) is conserved upon receptor binding. An excessive restriction in the motions of the aromatic side chain, imposed by the highly steric constraint of the indolizidine moiety, emerges as an alternative explanation. The findings reported here demonstrate the possibility of replacing the Pro¹⁰-Tyr¹¹ dipeptide in NT_8–13 with a non-peptide residue without affecting considerably the affinity for brain NT receptors.     

Skin graft rejection and delayed-type hypersensitivity responses to H-Y in an I-A b mutant

Based on graft rejection in C57BL/6 and B10.A(4R), but not in B10.A mice, skin graft rejection and delayed-type hypersensitivity (DTH) responses to the male HY antigen were considered to be under the control of the IBb gene in the mouse H-2 complex. These two phenomena were re-examined in the B6.C-H-2 ^bm12 mutant strain [mutation in the A gene in IA leading to an alteration in Ia ^b serologically detected specificities and the inability to generate cytotoxic T (Tc) cells to H-Y]. In this study the bm12 mutant was shown to produce weak DTH responses to H-Y. By contrast, bm12 female mice were unable to reject male skin grafts unless they had received prior footpad priming of male spleen cells, when graft rejection occurred, albeit slowly. In C57BL/6 mice the response to the HY antigen therefore appears to be solely under the control of the IA ^b gene. In other strains, response/nonresponse is presumably dictated by the ability of IA/IE interactions to produce T-helper responses.