Glycoprotein misfolding in the endoplasmic reticulum: identification of released oligosaccharides reveals a second ER-associated degradation pathway for Golgi-retrieved proteins

Endoplasmic reticulum-associated degradation (ERAD) is a key cellular process whereby misfolded proteins are removed from the endoplasmic reticulum (ER) for subsequent degradation by the ubiquitin/proteasome system. In the present work, analysis of the released, free oligosaccharides (FOS) derived from all glycoproteins undergoing ERAD, has allowed a global estimation of the mechanisms of this pathway rather than following model proteins through degradative routes. Examining the FOS produced in endomannosidase-compromised cells following α-glucosidase inhibition has revealed a mechanism for clearing Golgi-retrieved glycoproteins that have failed to enter the ER quality control cycle. The Glc₃Man₇GlcNAc₂ FOS species has been shown to be produced in the ER lumen by a mechanism involving a peptide: N-glycanase-like activity, and its production was sensitive to disruption of Golgi-ER trafficking. The detection of this oligosaccharide was unaffected by the overexpression of EDEM1 or cytosolic mannosidase, both of which increased the production of previously characterised cytosolically localised FOS. The lumenal FOS identified are therefore distinct in their production and regulation compared to FOS produced by the conventional route of misfolded glycoproteins directly removed from the ER. The production of such lumenal FOS is indicative of a novel degradative route for cellular glycoproteins that may exist under certain conditions.     

Impacts of black-tailed prarie dog rodenticides on nontarget passerines

In 1983 zinc phosphide, strychnine with prebait, and strychnine without prebait were applied to black-tailed prairie dog ( Cynomys ludovicianus ) colonies in west central South Dakota. Short-term (four days later) and long-term (one year later) impacts of the rodenticides on Horned Larks ( Eremophila alpestris ) and other granivorous birds inhabiting prairie dog colonies were evaluated. Hrned Larks and 49 other bird species were observed. Immediate impacts reduced Horned Lark relative densities 66% with strychnine only and 55% with prebaited strychnine. Zinc phosphide caused no measurable reduction. Horned Larks showed no long-term direct impacts. Indirect negative impacts occurred through habitat changes following prairie dog control. The granivorous guild showed no short- or long-term effects.     

Dry-year grazing and Nebraska sedge ( Carex nebraskensis )

In 1984 (a dry year), Tule Meadow, in the Sierra National Forest, California, was well grazed after several years of light use. This situation provided the opportunity to study responses of Nebraska sedge ( Carex nebraskensis ), an important forage species in mountain meadows, to protection and grazing. Rooted shoot frequencies and densities in fall 1984 and spring 1985 were the same within an exclosure and on the grazed area. Residual herbage (shoot weight) in fall and shoot heights in spring were greater within the exclosure. Lower spring shoot heights on the grazed area may relate to fall regrowth and reduced insulation induced by grazing. Nitrogen and potassium content of fall herbage was greater on the grazed area. Phosphorus content was the same both inside and outside the exclosure.     

Structure of the nociceptin/orphanin FQ receptor in complex with a peptide mimetic

Members of the opioid receptor family of G-protein-coupled receptors (GPCRs) are found throughout the peripheral and central nervous system, where they have key roles in nociception and analgesia. Unlike the 'classical' opioid receptors, δ, κ and μ (δ-OR, κ-OR and μ-OR), which were delineated by pharmacological criteria in the 1970s and 1980s, the nociceptin/orphanin FQ (N/OFQ) peptide receptor (NOP, also known as ORL-1) was discovered relatively recently by molecular cloning and characterization of an orphan GPCR. Although it shares high sequence similarity with classical opioid GPCR subtypes (～60%), NOP has a markedly distinct pharmacology, featuring activation by the endogenous peptide N/OFQ, and unique selectivity for exogenous ligands. Here we report the crystal structure of human NOP, solved in complex with the peptide mimetic antagonist compound-24 (C-24) (ref. 4), revealing atomic details of ligand-receptor recognition and selectivity. Compound-24 mimics the first four amino-terminal residues of the NOP-selective peptide antagonist UFP-101, a close derivative of N/OFQ, and provides important clues to the binding of these peptides. The X-ray structure also shows substantial conformational differences in the pocket regions between NOP and the classical opioid receptors κ (ref. 5) and μ (ref. 6), and these are probably due to a small number of residues that vary between these receptors. The NOP-compound-24 structure explains the divergent selectivity profile of NOP and provides a new structural template for the design of NOP ligands.     

The genomic clone G-21 which resembles a beta-adrenergic receptor sequence encodes the 5-HT1A receptor

The recent cloning of the complementary DNAs and/or genes for several receptors linked to guanine nucleotide regulatory proteins including the adrenergic receptors (alpha 1, alpha 2A, alpha 2B, beta 1, beta 2), several subtypes of the muscarinic cholinergic receptors, and the visual 'receptor' rhodopsin has revealed considerable similarity in the primary structure of these proteins. In addition, all of these proteins contain seven putative transmembrane alpha-helices. We have previously described a genomic clone, G-21, isolated by cross-hybridization at reduced stringency with a full length beta 2-adrenergic receptor probe. This clone contains an intronless gene which, because of its striking sequence resemblance to the adrenergic receptors, is presumed to encode a G-protein-coupled receptor. Previous attempts to identify this putative receptor by expression studies have failed. We now report that the protein product of the genomic clone, G21, transiently expressed in monkey kidney cells has all the typical ligand-binding characteristics of the 5-hydroxytryptamine (5-HT1A) receptor.     

Rangeland alpha diversities: Harvey Valley, Lassen National Forest, California

Nutrients in soil covered by Carex exserta sod and in adjacent unvegetated gravel areas were compared at Siberian Outpost, Sequoia National Park, California. The comparisons were part of a study to learn if Carex exserta meadow can be reestablished and if herbaceous cover on gravel areas can be increased. Grazing capacity and aesthetic appeal of denuded areas would be improved by better vegetative cover. The sod had higher concentrations of calcium, copper, iron, magnesium, manganese, nitrogen, potassium, and zinc than did the gravel areas. And it had a higher soil pH and percent organic matter. Sod and gravel did not differ in concentrations of phosphorus and sulfur. The differences were as might be expected between climax and badly deteriorated (or early seral) situations, and the results suggest that fertilization may be a useful treatment.         

Erythrocyte catechol-O-methyltransferase and plasma dopamine-β-hydroxylase in human umbilical cord blood

Summary Plasma dopamine--hydroxylase enzymatic activity and immunoreactive protein levels in human umbilical cord blood are only about 2% as great as values in the blood of older subjects. Erythrocyte catechol-O-methyltransferase activity levels in umbilical cord blood are very similar to those in the blood of adult subjects.This work was supported in part by NIH grants NS 11014 and HL 17487-1. Dr Weinshilboum is an Established Investigator of the American Heart Association. We thank Luanne Wussow and the physicians and nurses of the Department of Obstetrics and Gynecology of the Mayo Clinic for their assitance with these studies.