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The formation and the chemical characterization of single atoms of dubnium (Db, element 105), in the form of its volatile oxychloride, was investigated using the on-line gas phase chromatography technique, in the temperature range 350-600 °C. Under the exactly same chemical conditions, comparative studies with the lighter homologues of Groupâ 5 in the Periodic Table clearly indicate the volatility sequence being NbOCl3 > TaOCl3 ≥ DbOCl3 . From the obtained experimental results, thermochemical data for DbOCl3 were derived. The present study delivers reliable experimental information for theoretical calculations on chemical properties of transactinides.
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Here, we present a review on a fundamental radiochemical research topic performed by Swiss scientists in national and international collaborations, utilizing large accelerator facilities at the Paul Scherrer Institute as well as abroad. The chemical investigation of the heaviest elements of the periodic table is a truly multidisciplinary effort, which allows scientists to venture into a variety of fields ranging from nuclear and radiochemistry to experimental and theoretical work in inorganic and physical chemistry all the way to nuclear and atomic physics. The structure and fundamental ordering scheme of all elements in the periodic table, as established more than 150 years ago, is at stake: The ever increasing addition of new elements at the heavy end of the periodic table together with a growing influence of relativistic effects, raises the question of how much periodicity applies in this region of the table. Research on the heaviest chemical elements requires access to large heavy-ion accelerator facilities as well as to rare actinide isotopes as target materials. Thus, this scientific area is inevitably embedded in joint international efforts. Its fundamental character ensures academic relevance and thereby substantially contributes to the future of nuclear sciences in Switzerland.
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A gas-phase chemical study of rhenium carbonyls was carried out using short-lived radioisotopes produced at a heavy-ion accelerator. The Re isotopes produced in the nuclear reactions of natGd(23Na,xn)172-177Re were pre-separated with a gas-filled recoil ion separator and their carbonyls were synthesized in a mixture of inert gas and carbon monoxide. Using a low temperature isothermal chromatography apparatus, the adsorption enthalpies of Re carbonyls were derived to be ΔHads = -42 ± 2 kJ mol-1 on a Teflon® surface by fitting the external chromatograms with a Monte Carlo simulation program. A chemical yield of 25% relative to that of the transport yield for Re by a He/KCl gas-jet was achieved. The laser-ablation time-of-flight mass-spectrometric technique was employed to identify the species of Re carbonyls produced in the gas phase. The most stable species was deduced to be Re(CO)5 based on the mass-spectrometric analysis as well as quantum chemistry calculations.
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Interactions of single atoms of element 120 (E120) and its lighter homologs (Ba and Ra) with the stable gold (111) surface simulated by clusters are studied using relativistic density functional theory and accurate two-component shape-consistent small-core pseudopotentials. The predicted E120 adsorption energy on gold (ca. 250 kJ mol(-1)) is significantly larger than the previously reported value. The trends in interactions of heavy group 2 elements with gold are discussed on the basis of electronic structure calculations and estimates by the semiempirical macroscopic Eichler-Miedema model.
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Introduction: Targeted Radionuclide Therapy is used for the treatment of tumors in nuclear medicine, while sparing healthy tissues. Its application to cancer treatment is expanding. In particular, Auger-electron emitters potentially exhibit high efficacy in treating either small metastases or single tumor cells due to their short range in tissue. The aim of this paper is to study the feasibility of a large-scale production of thulium-167, an Auger-electron emitter radionuclide, in view of eventual systematic preclinical studies. Methods: Proton-irradiated enriched erbium-167 and erbium-168 oxides were used to measure the production cross sections of thulium-165, thulium-166, thulium-167, and thulium-168 utilizing an 18-MeV medical cyclotron equipped with a Beam Transport Line (BTL) at the Bern medical cyclotron laboratory. The comparison between the experimental and the TENDL 2021 theoretical cross-section results were in good agreement. Additional experiments were performed to assess the production yields of thulium radioisotopes in the BTL. Thulium-167 production yield was also measured irradiating five different target materials (167 Er 2 O 3, 168 Er 2 O 3, nat Tm 2 O 3, nat Yb 2 O 3, 171 Yb 2 O 3) with proton beams up to 63 MeV at the Injector II cyclotron of Paul Scherrer Institute. Results and Discussion: Our experiments showed that an 8-h irradiation of enriched ytterbium-171 oxide produced about 420 MBq of thulium-167 with a radionuclidic purity of 99.95% after 5 days of cooling time with a proton beam of about 53 MeV. Larger activities of thulium-167 can be achieved using enriched erbium-168 oxide with a 23-MeV proton beam, obtaining about 1 GBq after 8-h irradiation with a radionuclidic purity of <99.5% 5 days post end of bombardment.
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Carbonyl complexes of radioactive transition metals can be easily synthesized with high yields by stopping nuclear fission or fusion products in a gas volume containing CO. Here, we focus on Mo, W, and Os complexes. The reaction takes place at pressures of around 1 bar at room temperature, i.e., at conditions that are easy to accommodate. The formed complexes are highly volatile. They can thus be transported within a gas stream without major losses to setups for their further investigation or direct use. The rapid synthesis holds promise for radiochemical purposes and will be useful for studying, e.g., chemical properties of superheavy elements.
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We report on the application of an innovative whey protein amyloids-carbon hybrid filter for the removal and disposal of the long-lived radioactive fission product 137Cs from aqueous samples. Test experiments revealed a reduction of 137Cs radioactivity by a factor of 340 compared to the initial solution, with an efficiency as high as 99.7%. The adsorption capacity of the membrane was explored by performing several cycles of filtration, indicating a potential retention of more than 115 MBq per gram of filtering material at the applied experimental conditions. These results pave the way for further investigations on the applicability of this filter material to other nuclear fission products.
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Accelerator driven fast nuclear reactors cooled by lead-bismuth eutectic (LBE) are developed for transmuting long-lived radionuclides in spent nuclear fuel. Due to the nature of the coolant, operating the reactor will result in a production of 210Po by neutron capture. Understanding the behavior of this highly radiotoxic nuclide in the event of a failure of the window separating the evacuated proton beam guide from the reactor core is required for safety assessments. The present work aims at acquiring this knowledge by studying the evaporation of polonium from neutron-irradiated LBE and its deposition in a scaled down model of the beam tube. Experimental results along with Monte Carlo simulations indicate that polonium adsorbs as a single species with an adsorption enthalpy of approximately -156 kJ/mol.
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BACKGROUND: Rubella infections are usually characterized by mild self-limiting courses in immunocompetent individuals. However, infections in pregnant women during the first trimester of pregnancy pose a high risk of congenital rubella syndrome possibly resulting in severe defects in the unborn child. Rubella serology of a primary rubella infection is mainly determined by diagnostic confirmation of levels of specific IgM. STUDY DESIGN: Here, we report on the performance of the Rubella IgM assay in development on the ARCHITECT instrument, a fully automated high throughput chemiluminescent microparticle immunoassay platform. Sensitivity was examined using commercially available seroconversion panels from vaccinated individuals; specificity was addressed by testing populations of pregnant women, blood donors and hospitalized patients. In addition, the potential for assay interference was evaluated by testing samples of several disease states. As methods of comparison AxSYM, BioMérieux VIDAS and Behring Rubella IgM assays were used. RESULTS: The study demonstrates that the ARCHITECT Rubella IgM assay shows improved specificity compared to AxSYM and Behring. Seroconversion sensitivity is equivalent on all assays evaluated. CONCLUSION: Together with high throughput, optimized specificity and suppression of rheumatoid factor (RF) interference the ARCHITECT assay provides a useful improvement for the diagnosis of rubella serology.
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Anticorpos Antivirais/sangue , Imunoglobulina M/sangue , Complicações Infecciosas na Gravidez/diagnóstico , Kit de Reagentes para Diagnóstico , Vírus da Rubéola/imunologia , Rubéola (Sarampo Alemão)/diagnóstico , Automação , Doadores de Sangue , Feminino , Humanos , Imunoensaio/métodos , Gravidez , Complicações Infecciosas na Gravidez/virologia , Rubéola (Sarampo Alemão)/virologia , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: The favorable decay properties of 43Sc and 44Sc for PET make them promising candidates for future applications in nuclear medicine. An advantage 43Sc (T1/2 = 3.89 h, Eß+av = 476 keV [88%]) exhibits over 44Sc, however, is the absence of co-emitted high energy γ-rays. While the production and application of 44Sc has been comprehensively discussed, research concerning 43Sc is still in its infancy. This study aimed at developing two different production routes for 43Sc, based on proton irradiation of enriched 46Ti and 43Ca target material. RESULTS: 43Sc was produced via the 46Ti(p,α)43Sc and 43Ca(p,n)43Sc nuclear reactions, yielding activities of up to 225 MBq and 480 MBq, respectively. 43Sc was chemically separated from enriched metallic 46Ti (97.0%) and 43CaCO3 (57.9%) targets, using extraction chromatography. In both cases, ~90% of the final activity was eluted in a small volume of 700 µL, thereby, making it suitable for direct radiolabeling. The prepared products were of high radionuclidic purity, i.e. 98.2% 43Sc were achieved from the irradiation of 46Ti, whereas the product isolated from irradiated 43Ca consisted of 66.2% 43Sc and 33.3% 44Sc. A PET phantom study performed with 43Sc, via both nuclear reactions, revealed slightly improved resolution over 44Sc. In order to assess the chemical purity of the separated 43Sc, radiolabeling experiments were performed with DOTANOC, attaining specific activities of 5-8 MBq/nmol, respectively, with a radiochemical yield of >96%. CONCLUSIONS: It was determined that higher 43Sc activities were accessible via the 43Ca production route, with a comparatively less complex target preparation and separation procedure. The product isolated from irradiated 46Ti, however, revealed purer 43Sc with minor radionuclidic impurities. Based on the results obtained herein, the 43Ca route features some advantages (such as higher yields and direct usage of the purchased target material) over the 46Ti path when aiming at 43Sc production on a routine basis.
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Lassa virus glycoprotein is synthesised as a precursor (preGP-C) into the lumen of the endoplasmic reticulum. After cotranslational cleavage of the signal peptide, the immature GP-C is posttranslationally processed into the N-terminal subunit GP-1 and the C-terminal subunit GP-2 by the host cell subtilase SKI-1/S1P. The glycoprotein precursor contains eleven potential N-glycosylation sites. In this report, we investigated the effect of each N-glycan on proteolytic cleavage and cell surface transport by disrupting the consensus sequences of eleven potential N-glycan attachment sites individually. Five glycoprotein mutants with disrupted N-glycosylation sites were still proteolytically processed, whereas the remaining N-glycosylation sites are necessary for GP-C cleavage. Despite the lack of proteolytic processing, all cleavage-defective mutants were transported to the cell surface and remained completely endo H-sensitive. The findings indicate that N-glycans are needed for correct conformation of GP-C in order to be cleaved by SKI-1/S1P.
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Vírus Lassa/metabolismo , Polissacarídeos/metabolismo , Proteínas do Envelope Viral/metabolismo , Animais , Membrana Celular/metabolismo , Chlorocebus aethiops , Glicoproteínas/genética , Glicoproteínas/metabolismo , Glicosilação , Vírus Lassa/genética , Mutação , Pró-Proteína Convertases/metabolismo , Conformação Proteica , Dobramento de Proteína , Precursores de Proteínas/genética , Precursores de Proteínas/metabolismo , Serina Endopeptidases/metabolismo , Transfecção , Células Vero , Proteínas do Envelope Viral/química , Proteínas do Envelope Viral/genéticaRESUMO
The Z protein is the matrix protein of arenaviruses and has been identified as the main driving force for budding. Both LCMV and Lassa virus Z proteins bud from cells in the absence of other viral proteins as enveloped virus-like particles. Z accumulates near the inner surface of the plasma membrane where budding takes place. Furthermore, biochemical data have shown that Z is strongly membrane associated. The primary sequence of Z lacks a typical transmembrane domain and until now it is not understood by which mechanism Z is able to interact with cellular membranes. In this report, we analyzed the role of N-terminal myristoylation for the membrane binding of Lassa virus Z. We show that disruption of the N-terminal myristoylation signal by substituting the N-terminal glycine with alanine (Z-G2A mutant) resulted in a significant reduction of Z protein association with cellular membranes. Furthermore, removal of the myristoylation site resulted in a relocalization of Z from a punctuate distribution to a more diffuse cellular distribution pattern. Finally, treatment of Lassa virus-infected cells with various myristoylation inhibitors drastically reduced efficient Lassa virus replication. Our data indicate that myristoylation of Z is critical for its binding ability to lipid membranes and thus, for effective virus budding.
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Proteínas de Transporte/metabolismo , Membrana Celular/metabolismo , Vírus Lassa/metabolismo , Ácido Mirístico/metabolismo , Processamento de Proteína Pós-Traducional , Proteínas da Matriz Viral/metabolismo , Sequência de Aminoácidos , Animais , Sítios de Ligação , Proteínas de Transporte/química , Proteínas de Transporte/genética , Linhagem Celular Tumoral , Chlorocebus aethiops , Humanos , Mutação , Ligação Proteica , Transporte Proteico , Proteínas de Ligação a RNA , Células Vero , Proteínas da Matriz Viral/química , Proteínas da Matriz Viral/genéticaRESUMO
During the last decade, a number of related bacterial membrane-bound multihaem c-type cytochromes, collectively referred to as the NapC/NirT family, were identified. These proteins are generally thought to catalyse electron transport between the quinone/quinol pool and periplasmic oxidoreductases. The best-characterized members, the tetrahaem c-type cytochromes NrfH and NapC, mediate electron transport to NrfA and NapA respectively. Amino acid sequence alignments suggest that the nature and position of distal haem c iron ligands differs in NrfH and NapC proteins. Site-directed modification of potential haem c iron-ligating histidine, lysine and methionine residues in Wolinella succinogenes NrfH was performed to determine the implication in electron transport from formate to nitrite. Two histidine, one lysine and one methionine residues were found to be essential, whereas the replacement of three other conserved histidine residues, one methionine and two lysines did not prevent growth by nitrite respiration. The results contrast those previously obtained for Paracoccus pantotrophus NapC, in which four essential histidine residues have been identified that are highly likely to serve as distal haem c iron ligands. The combined experimental evidence suggests different haem ligation patterns within NapC and NrfH proteins, which might reflect their different functions in the bacterial electron transfer.
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Grupo dos Citocromos c/genética , Grupo dos Citocromos c/metabolismo , Heme/análogos & derivados , Ferro/metabolismo , Mutagênese Sítio-Dirigida , Wolinella/enzimologia , Wolinella/genética , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Grupo dos Citocromos c/química , Genes Bacterianos/genética , Heme/química , Heme/metabolismo , Ferro/química , Mutação , Nitratos/metabolismo , Nitritos/metabolismo , Wolinella/metabolismoRESUMO
Lassa virus glycoprotein is synthesized as precursor GP-C into the lumen of the endoplasmic reticulum and cleaved posttranslationally into the N-terminal subunit GP-1 and the C-terminal subunit GP-2 by subtilase SKI-1/S1P. The N-terminal portion of the primary translation product preGP-C contains a signal peptide of unknown length. In order to demonstrate the signal peptide cleavage site, purified viral GP-1 isolated from Lassa virus particles was N-terminally sequenced as TSLYKGV, identical to amino acids 59-65 of GP-C. Mutational analysis of the amino acid residues flanking the putative cleavage site led to non-cleavable preGP-C indicating that no other signal peptide cleavage site exists. Interestingly, GP-C mutants with a non-cleavable signal peptide were not further processed by SKI-1/S1P. This observation suggests that the signal peptide cleavage is necessary for GP-C maturation and hence for Lassa virus replication.
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Glicoproteínas/metabolismo , Vírus Lassa/metabolismo , Sinais Direcionadores de Proteínas , Proteínas Virais/metabolismo , Animais , Chlorocebus aethiops , Eletroforese em Gel de Poliacrilamida , Glicoproteínas/química , Hidrólise , Células Vero , Proteínas Virais/químicaRESUMO
The two multiheme c-type cytochromes NrfH and NrfA form a membrane-bound complex that catalyzes menaquinol oxidation by nitrite during respiratory nitrite ammonification of Wolinella succinogenes. Each cysteine residue of the four NrfH heme c binding motifs was individually replaced by serine. Of the resulting eight W. succinogenes mutants, only one is able to grow by nitrite respiration although its electron transport activity from formate to nitrite is decreased. NrfH from this mutant was shown by matrix-assisted laser desorption/ionization mass spectrometry to carry four covalently bound heme groups like wild-type NrfH indicating that the cytochrome c biogenesis system II organism W. succinogenes is able to attach heme to an SXXCH motif.
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Grupo dos Citocromos c/metabolismo , Citocromos a1 , Citocromos c1 , Heme/análogos & derivados , Heme/metabolismo , Nitrato Redutases/metabolismo , Proteínas de Ligação a RNA , Wolinella/metabolismo , Motivos de Aminoácidos , Proteínas de Bactérias/metabolismo , Sítios de Ligação , Meios de Cultura , Cisteína/genética , Cisteína/metabolismo , Mutagênese Sítio-Dirigida , Nitrato Redutases/genética , Nitritos/metabolismo , Oxirredução , Serina/genética , Serina/metabolismo , Fatores de Transcrição/metabolismo , Wolinella/genética , Wolinella/crescimento & desenvolvimentoRESUMO
Lassa virus is the causative agent of a hemorrhagic fever endemic in west Africa. The RNA genome of Lassa virus encodes the glycoprotein precursor GP-C, a nucleoprotein (NP), the viral polymerase L and a small protein Z (11 kDa). Here, we analyze the role of Z protein for virus maturation. We have recently shown that expression of Z protein in the absence of other viral proteins is sufficient for the release of enveloped Z-containing particles. In this study, we examined particles secreted into the supernatant of a stably Z protein-expressing CHO cell line by electron microscopy. The observed Z-induced virus-like particles did not significantly differ in their morphology and size from Lassa virus particles. Mutation of two proline-rich domains within Z which are known to drastically reduce the release of virus-like particles, had no effect on the cellular localization of the protein nor on its membrane-association. Furthermore, we present evidence that Z interacts with the NP. We assume that Z recruits NP to cellular membranes where virus assembly takes place. We conclude from our data that Lassa virus Z protein plays an essential role in Lassa virus maturation.
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Proteínas de Transporte/genética , Proteínas de Transporte/ultraestrutura , Vírus Lassa/ultraestrutura , Nucleoproteínas/metabolismo , Proteínas da Matriz Viral/genética , Proteínas da Matriz Viral/ultraestrutura , Sequência de Aminoácidos , Substituição de Aminoácidos , Arenavirus/genética , Sítios de Ligação , Proteínas de Transporte/metabolismo , Genoma Viral , Vírus Lassa/genética , Dados de Sequência Molecular , Mutação , Nucleoproteínas/ultraestrutura , RNA Viral/genética , Proteínas de Ligação a RNA , Alinhamento de Sequência , Deleção de Sequência , Homologia de Sequência de Aminoácidos , Proteínas da Matriz Viral/metabolismoRESUMO
Lassa virus glycoprotein is translated as a precursor (pre-GP-C) into the lumen of the endoplasmic reticulum and is cotranslationally cleaved into the signal peptide and GP-C, before GP-C is proteolytically processed into its subunits GP1 and GP2. The signal peptide of pre-GP-C comprises 58 amino acids. The substitution of Lassa virus pre-GP-C signal peptide with another signal peptide still mediates translocation and the release of signal peptide but abolishes the proteolytic cleavage of GP-C into GP1 and GP2. Remarkably, cleavage of GP-C from these hybrid pre-GP-C substrates was restored on coexpression of the wild-type pre-GP-C signal peptide, indicating that the signal peptide functions as a trans-acting factor to promote Lassa virus GP-C processing. To our knowledge, this is the first report on a signal peptide that is essential for proteolytic processing of a secretory pathway protein.
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Vírus Lassa/metabolismo , Precursores de Proteínas/metabolismo , Sinais Direcionadores de Proteínas/fisiologia , Transativadores/metabolismo , Processamento de Proteína Pós-Traducional/fisiologia , Proteínas do Envelope Viral/metabolismoRESUMO
The cytochrome c nitrite reductase complex (NrfHA) is the terminal enzyme in the electron transport chain catalysing nitrite respiration of Wolinella succinogenes. The catalytic subunit NrfA is a pentahaem cytochrome c containing an active site haem group which is covalently bound via the cysteine residues of a unique CWTCK motif. The lysine residue serves as the axial ligand of the haem iron. The other four haem groups of NrfA are bound by conventional haem-binding motifs (CXXCH). The nrfHAIJ locus was restored on the genome of the W. succinogenes DeltanrfAIJ deletion mutant by integration of a plasmid, thus enabling the expression of modified alleles of nrfA and nrfI. A mutant (K134H) was constructed which contained a nrfA gene encoding a CWTCH motif instead of CWTCK. NrfA of strain K134H was found to be synthesized with five bound haem groups, as judged by matrix-assisted laser-desorption/ionization (MALDI) mass spectrometry. Its nitrite reduction activity with reduced benzyl viologen was 40% of the wild-type activity. Ammonia was formed as the only product of nitrite reduction. The mutant did not grow by nitrite respiration and its electron transport activity from formate to nitrite was 5% of that of the wild-type strain. The predicted nrfI gene product is similar to proteins involved in system II cytochrome c biogenesis. A mutant of W. succinogenes (stopI) was constructed that contained a nrfHAIJ gene cluster with the nrfI codons 47 and 48 altered to stop codons. The NrfA protein of this mutant did not catalyse nitrite reduction and lacked the active site haem group, whereas the other four haem groups were present. Mutant (K134H/stopI) which contained the K134H modification in NrfA in addition to the inactivated nrfI gene had essentially the same properties as strain K134H. NrfA from strain K134H/stopI contained five haem groups. It is concluded that NrfI is involved in haem attachment to the CWTCK motif in NrfA but not to any of the CXXCH motifs. The nrfI gene is obviously dispensable for maturation of a modified NrfA protein containing a CWTCH motif instead of CWTCK. Therefore, NrfI might function as a specific haem lyase that recognizes the active site lysine residue of NrfA. A similar role has been proposed for NrfE, F and G of Escherichia coli, although these proteins share no overall sequence similarity to NrfI and belong to system I cytochrome c biogenesis, which differs fundamentally from system II.
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Proteínas de Bactérias/genética , Citocromos a1 , Citocromos c1 , Nitrato Redutases/metabolismo , Wolinella/metabolismo , Substituição de Aminoácidos , Proteínas de Bactérias/metabolismo , Histidina , Lisina , Nitrato Redutases/genética , Wolinella/genéticaRESUMO
Lassa virus glycoprotein C (GP-C) is translated as a precursor (preGP-C) into the lumen of the endoplasmic reticulum (ER) and cotranslationally cleaved into the signal peptide and immature GP-C before GP-C is proteolytically processed into its subunits, GP-1 and GP-2, which form the mature virion spikes. The signal peptide of preGP-C comprises 58 amino acids and contains two distinct hydrophobic domains. Here, we show that each hydrophobic domain alone can insert preGP-C into the ER membrane. Furthermore, we demonstrate that the native signal peptide only uses the N-terminal hydrophobic domain for membrane insertion, exhibiting a novel type of a topology for signal peptides with an extended ER luminal part, which is essential for proteolytic processing of GP-C into GP-1 and GP-2.