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Affinity precipitation and site-specific immobilization of proteins carrying polyhistidine tails

Proteins carrying genetically attached polyhistidine tails have been purified using affinity precipitation with metal chelates. DNA fragments encoding four or five histidine residues have been genetically fused to the oligomeric enzymes lactate dehydrogenase (Bacillus stearothermophilus), beta-glucoronidase (Escherichia coli), and galactose dehydrogenase (Pseudomonas fluorescens) as well as to the

Site-directed primary in vitro immunization : Production of HIV-1 neutralizing human monoclonal antibodies from lymphocytes obtained from seronegative donors

The design of an in vitro immunization system based on a synthetic heterotope immunogen, which was a peptide containing both T- and B-cell epitopes, that elicited a neutralizing, primary human humoral immune response against the human immunodeficiency virus (HIV-1) is reported here. This heterotope construct contained the major neutralizing B-cell epitope, within the V3 region of glycoprotein 120

A framework for analyzing deployment of solar photovoltaics, with a focus on building-sited grid-connected systems

Policy intervention schemes for increased deployment of solar photovoltaics (PV) have been launched in several countries, with varying success. In order to ensure an efficient deployment of the technology, thorough knowledge is needed about relevant present actors and institutions, and about how a desirable actor base and institutional setup should look like; mere cost reductions are not enough to

Localization of thrombin cleavage sites in the amino-terminal region of bovine protein S

Protein S is a vitamin K-dependent plasma protein. It functions as a cofactor to activated protein C in the inactivation of factors Va and VIIIa by limited proteolysis. Protein S is very sensitive to proteolysis by thrombin which reduces its calcium ion binding and leads to a loss of its cofactor activity. We have now determined the sequence of the 100 amino-terminal amino acid residues and locali

Intercomparison of eddy-covariance software for urban tall-tower sites

Long-term tall-tower eddy-covariance (EC) measurements have been recently established in three European pilot cities as part of the ICOS-Cities project. We conducted a comparison of EC software to ensure a reliable generation of interoperable flux estimates, which is the prerequisite for avoiding methodological biases and improving the comparability of the results. We analyzed datasets covering 5

Two-site recognition of Staphylococcus aureus peptidoglycan by lysostaphin SH3b

Lysostaphin is a bacteriolytic enzyme targeting peptidoglycan, the essential component of the bacterial cell envelope. It displays a very potent and specific activity toward staphylococci, including methicillin-resistant Staphylococcus aureus. Lysostaphin causes rapid cell lysis and disrupts biofilms, and is therefore a therapeutic agent of choice to eradicate staphylococcal infections. The C-term

Self-replication of Aβ42 aggregates occurs on small and isolated fibril sites

Self-replication of amyloid fibrils via secondary nucleation is an intriguing physicochemical phenomenon in which existing fibrils catalyze the formation of their own copies. The molecular events behind this fibril surface-mediated process remain largely inaccessible to current structural and imaging techniques. Using statistical mechanics, computer modeling, and chemical kinetics, we show that th

Comparison of tTEM-IP and ERT-IP : cases from mine tailing sites in Sweden

The induced polarization (IP) effect can be studied using both galvanic and inductive methods. Here we study IP and its manifestation in transient electromagnetic (TEM) data from a tTEM system and in time-domain electric resistivity tomography (ERT) data. Since the two methods are sensitive to two different frequency ranges, a direct IP spectrum comparison is not possible. Instead, we assess the I

The declining spadefoot toad Pelobates fuscus: calling site choice and conservation

We investigated whether local biotic and abiotic conditions could explain the occurrence of calling males of the declining spadefoot toad Pelobates fuscus in 72 ponds in southern Sweden. The ponds covered the entire distribution range of P. fuscus and were monitored during the breeding season in 2000. Calling males were found in 33 ponds. representing ca 50% of all known ponds for the species ill

Structure-Based Dissection of the Active Site Chemistry of Leukotriene A4 Hydrolase: Implications for M1 Aminopeptidases and Inhibitor Design

M1 aminopeptidases comprise a large family of biologically important zinc enzymes. We show that peptide turnover by the M1 prototype, leukotriene A4 hydrolase/aminopeptidase, involves a shift in substrate position associated with exchange of zinc coordinating groups, while maintaining the overall coordination geometry. The transition state is stabilized by residues conserved among M1 members and i

Calbindin D-28k EF-hand ligand binding and oligomerization: Four high-affinity sites-three modes of action

Calbindin D-28k, a highly conserved protein with Ca2+-sensing and Ca2+-buffering capabilities, is abundant in brain and sensory neurons. This protein contains six EF-hand subdomains, four of which bind Call with high affinity. Calbindin D28k can be reconstituted from six synthetic peptides corresponding to the six EF-hands, indicating a single-domain structure with multiple interactions between th

Modulation of GABA(A) Receptors by Natural Products and the Development of Novel Synthetic Ligands for the Benzodiazepine Binding Site.

Nature provides science and society with a virtually unlimited supply of structurally diverse and biologically active molecules; the natural products. While some are directly useful in commercial applications, others are valuable for studying and understanding biological phenomena at the molecular level. An example is the signaling of nerve cells, which has been explored in considerable detail usi

Pedon-scale silicate weathering : comparison of the PROFILE model and the depletion method at 16 forest sites in Sweden

Weathering of soil minerals is important for the recovery from acidification and for the sustainability of forestry. However, there is still substantial uncertainty about its absolute rate. This study presents a harmonized comparison of field weathering rates estimated with the mechanistic model PROFILE and the depletion method for 16 intensively sampled soil profiles across Sweden representing di

Spatial and temporal dynamics in eddy covariance observations of methane fluxes at a tundra site in northeastern Siberia

In the past two decades, the eddy covariance technique has been used for an increasing number of methane flux studies at an ecosystem scale. Previously, most of these studies used a closed path setup with a tunable diode laser spectrometer (TDL). Although this method worked well, the TDL has to be calibrated regularly and cooled with liquid nitrogen or a cryogenic system, which limits its use in r

Micro-climate determines oviposition site selection and abundance in the butterfly Pyrgus armoricanus at its northern range margin

1. Knowledge about species habitat requirements is important when designing conservation strategies as well as for predicting species distributions. For herbivorous insects, insights in oviposition preferences can provide important information on their habitat requirements. 2. The oviposition preferences of Oberthür's Grizzled Skipper butterfly Pyrgus armoricanus Oberthür at its northern range bou