Biomedical subjects
C M Carr
Publications and source records attributed to C M Carr.
Ordering the final events in yeast exocytosis.
In yeast, assembly of exocytic soluble N-ethylmaleimide-sensitive fusion protein (NSF) attachment protein receptor (SNARE) complexes between the secretory vesicle SNARE Sncp and the plasma membrane SNAREs Ssop and Sec9p occurs at a late stage of the exocytic reaction. Mutations that block either secretory vesicle delivery or tethering prevent SNARE complex assembly and the localization of Sec1p, a SNARE complex binding protein, to sites of secretion. By contrast, wild-type levels of SNARE complexes persist in the sec1-1 mutant after a secretory block is imposed, suggesting a role for Sec1p after SNARE complex assembly. In the sec18-1 mutant, cis-SNARE complexes containing surface-accessible Sncp accumulate in the plasma membrane. Thus, one function of Sec18p is to disassemble SNARE complexes on the postfusion membrane.
Membrane fusion. Changing partners.
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Sec1p binds to SNARE complexes and concentrates at sites of secretion.
Proteins of the Sec1 family have been shown to interact with target-membrane t-SNAREs that are homologous to the neuronal protein syntaxin. We demonstrate that yeast Sec1p coprecipitates not only the syntaxin homologue Ssop, but also the other two exocytic SNAREs (Sec9p and Sncp) in amounts and in proportions characteristic of SNARE complexes in yeast lysates. The interaction between Sec1p and Ssop is limited by the abundance of SNARE complexes present in sec mutants that are defective in either SNARE complex assembly or disassembly. Furthermore, the localization of green fluorescent protein (GFP)-tagged Sec1p coincides with sites of vesicle docking and fusion where SNARE complexes are believed to assemble and function. The proposal that SNARE complexes act as receptors for Sec1p is supported by the mislocalization of GFP-Sec1p in a mutant defective for SNARE complex assembly and by the robust localization of GFP-Sec1p in a mutant that fails to disassemble SNARE complexes. The results presented here place yeast Sec1p at the core of the exocytic fusion machinery, bound to SNARE complexes and localized to sites of secretion.
Influenza hemagglutinin is spring-loaded by a metastable native conformation.
Enveloped viruses enter cells by protein-mediated membrane fusion. For influenza virus, membrane fusion is regulated by the conformational state of the hemagglutinin (HA) protein, which switches from a native (nonfusogenic) structure to a fusion-active (fusogenic) conformation when exposed to the acidic environment of the cellular endosome. Here we demonstrate that destabilization of HA at neutral pH, with either heat or the denaturant urea, triggers a conformational change that is biochemically indistinguishable from the change triggered by low pH. In each case, the conformational change is coincident with induction of membrane-fusion activity, providing strong evidence that the fusogenic structure is formed. These results indicate that the native structure of HA is trapped in a metastable state and that the fusogenic conformation is released by destabilization of native structure. This strategy may be shared by other enveloped viruses, including those that enter the cell at neutral pH, and could have implications for understanding the membrane-fusion step of HIV infection.
Flu virus invasion: halfway there.
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Subdomain folding of the coiled coil leucine zipper from the bZIP transcriptional activator GCN4.
One popular model for protein folding, the framework model, postulates initial formation of secondary structure elements, which then assemble into the native conformation. However, short peptides that correspond to secondary structure elements in proteins are often only marginally stable in isolation. A 33-residue peptide (GCN4-p1) corresponding to the GCN4 leucine zipper folds as a parallel, two-stranded coiled coil [O'Shea, E.K., Klemm, J.D., Kim, P.S., & Alber, T.A. (1991) Science 254, 539-544]. Deletion of the first residue (Arg 1) results in local, N-terminal unfolding of the coiled coil, suggesting that a stable subdomain of GCN4-p1 can form. N- and C-terminal deletion studies result in a 23-residue peptide, corresponding to residues 8-30 of GCN4-p1, that folds as a parallel, two-stranded coil with substantial stability (the melting temperature of a 1 mM solution is 43 degrees C at pH 7). In contrast, a closely related 23-residue peptide (residues 11-33 of GCN4-p1) is predominantly unfolded, even at 0 degrees C, as observed previously for many isolated peptides of similar length. Thus, specific tertiary packing interactions between two short units of secondary structure can be energetically more important in stabilizing folded structure than secondary structure propensities. These results provide strong support for the notion that stable, cooperatively folded subdomains are the important determinants of protein folding.
A spring-loaded mechanism for the conformational change of influenza hemagglutinin.
Influenza hemagglutinin (HA) undergoes a conformational change that induces viral fusion with the cellular membrane. The structure of HA in the fusogenic state is unknown. We have identified a sequence in HA that has a high propensity for forming a coiled coil. Surprisingly, this sequence corresponds to a loop region in the X-ray structure of native HA: the loop is followed by a three-stranded, coiled-coil stem. We find that a 36 residue peptide (LOOP-36), comprising the loop region and the first part of the stem, forms a three-stranded coiled coil. This coiled coil is extended and stabilized in a longer peptide, corresponding to LOOP-36 plus the residues of a preceding, short alpha helix. These findings lead to a model for the fusogenic conformation of HA: the coiled-coil stem of the native state extends, relocating the hydrophobic fusion peptide, by 100 A, toward the target membrane.
Midazolam-droperidol premedication for cardiac surgery. A comparison with papaveretum and hyoscine.
A combination of midazolam and droperidol given intramuscularly was compared with papaveretum and hyoscine for premedication of patients about to undergo cardiac surgery. Midazolam and droperidol proved to be a very satisfactory combination, producing superior sedation and anxiolysis with good cardiovascular stability.
The automatic implantable cardioverter-defibrillator. Implications for anaesthetists.
This paper describes the anaesthetic management of a patient who had an automatic implantable cardioverter-defibrillator. The working principles of the device, the indications for its insertion and the postoperative complications, are discussed. An increasing number of cardioverter-defibrillators are being implanted in the UK. At least two general anaesthetics are required for each patient; one for implantation of the device and a second for testing its efficiency in terminating ventricular tachycardia and ventricular fibrillation. In future, the number of patients presenting for noncardiac surgical procedures is likely to increase, therefore every anaesthetist should be aware of the problems involved in management.
Hemoglobinopathies in a general and family practice setting.
The incidence of sickle cell trait varies among blacks in North America. Although the average hemoglobin (Hb)-AS gene frequency is 8% in the United States, a value of 13.4% was recorded in South Carolina. Preliminary studies during a two-day community sponsored health fair in Selma, Alabama, revealed a sickle cell gene frequency of 15.96%. This study also found an intermediate frequency of 12.4% in a family practice center that is accredited for a residency program. Through a screening program, sickle cell trait was detected, a rigorous training program for resident physicians in family practice was implemented, and proper counseling to patients with hemoglobinopathies was provided. In the authors' judgment, laboratory screening for sickle cell trait is a first step toward genetic counseling and medical practice by family physicians and general practitioners.
Propofol reduces seizure duration in patients having anaesthesia for electroconvulsive therapy.
Twenty-five patients received either methohexitone 1.0 mg kg-1 or propofol 1.3 mg kg-1 to induce anaesthesia during two separate electroconvulsive therapy (ECT) treatments. A forearm was isolated before administration of suxamethonium 0.5 mg kg-1, so that unmodified seizure duration could be measured. Bifrontotemporal electrodes were applied to administer a standard 3-s ECT shock. Median (quartile deviation) duration of seizure was reduced significantly after propofol (19.0 (9.0) s), compared with after methohexitone (33.0 (7.8) s). Therefore propofol may not be an appropriate anaesthetic for ECT because of its adverse effect on seizure duration.
Repeated general anaesthetics in the presence of a large undiagnosed left atrial myxoma.
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Sickle gene frequency in a southcentral Alabama family medical program.
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Non-Hodgkin's lymphoma presenting initially as a sleep-apnea syndrome.
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A business coalition overcomes its suburban isolation.
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Sexual problems seen in family practice.
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Time-of-flight secondary ion mass spectrometry (ToF-SIMS) analysis of the application of a cationic conditioner to "clean" hair.
In this study the applicability of the surface-sensitive ToF-SIMS technique to hair analysis and associated aqueous processing is evaluated. ToF-SIMS analysis of "as received" human hair indicates the presence of silicones, anionic surfactants, and cationic conditioners, from previous treatments, on the fiber surface. Cleaning of the hair with SLS or SLES results in adsorption of the surfactants onto the fiber surface. In particular, the more non-polar surfactant components have greater substantivity for the fiber surface, as indicated by the relative increase in their ToF-SIMS intensity. Application of the Incroquat Behenyl 18-MEA conditioner to both "virgin" and bleached hair results in the adsorption of the cationic C18, C20, C22, and C21 surfactant components onto the hair surface. The ToF-SIMS data indicate higher levels of conditioner on the bleached hair relative to the undamaged hair.