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J T Pedersen

Publications and source records attributed to J T Pedersen.

At least 19 recordsLinked to original sources
PubMed1996

Genetic algorithms for protein structure prediction.

Genetic algorithms are a general class of search methods that mimic natural gene-based optimization mechanisms. Mutation, cross-over and replication operations are performed on strings. When applied to structure prediction, each string describes a particular conformation of a protein molecule. There are many ways in which such search methods may be implemented. Recent results show potential for helping with protein structure prediction, but more data are needed before a complete assessment can be made.

Algorithms↗
PubMed1995

[Tuberculosis in low prevalence countries. Back to the future?].

The incidence of tuberculosis has declined in all European countries in this century but in several countries the decline has come to a stop. Microscopy a.m. Ziehl-Neelsen and culture are still the mainstays in the diagnosis of tuberculosis but new DNA-technics such as PCR and RFLP are increasingly used. The tuberculin test is hitherto the only practically feasible test for identification of subjects infected with M. tuberculosis. BCG is the most used vaccine in the world but the protective mechanisms are not fully understood and the efficacy controversial. Immunotherapy with M. vaccae may be used as an adjuvant to chemotherapy. Few cases of tuberculosis are observed in AIDS-patients in Denmark. Modern standard treatment of tuberculosis comprises a six-month regimen with isoniazid, rifampicin, pyrazinamide and possibly ethambutol. Multidrug resistance in tuberculosis is rare in Western Europe. Chemoprophylaxis with isoniazid is debatable. Patients with smear-positive sputum may be infectious whereas patients with extrapulmonary tuberculosis normally pose no risk.

Disease Outbreaks↗
PubMed1995

Confronting the problem of interconnected structural changes in the comparative modeling of proteins.

Comparative models of three proteins have been built using a variety of computational methods, heavily supplemented by visual inspection. We consider the accuracy obtained to be worse than expected. A careful analysis of the models shows that a major reason for the poor results is the interconnectedness of the structural differences between the target proteins and the template structures they were modeled from. Side chain conformations are often determined by details of the structure remote in the sequence, and can be influenced by relatively small main chain changes. Almost all of the regions of substantial main chain conformational change interact with at least one other such region, so that they often cannot be modeled independently. Visual inspection is sometimes effective in correcting errors in sequence alignment and in spotting when an alternative template structure is more appropriate. We expect some improvements in the near future through the development of structure-based sequence alignment tools, side chain interconnectedness rotamer choice algorithms, and a better understanding of the context sensitivity of conformational features.

Amino Acid Sequence↗
PubMed1995

Ab initio structure prediction for small polypeptides and protein fragments using genetic algorithms.

Ab initio folding simulations have been performed on three peptides, using a genetic algorithm-based search method which operates on a full atom representation. Conformations are evaluated with an empirical force field parameterized by a potential of mean force analysis of experimental structures. The dominant terms in the force field are local and nonlocal main chain electrostatics and the hydrophobic effect. Two of the simulated structures were for fragments of complete proteins (eosinophil-derived neurotoxin (EDN) and the subtilisin propeptide) that were identified as being likely initiation sites for folding. The experimental structure of one of these (EDN) was subsequently found to be consistent with that prediction (using local hydrophobic burial as the determinant for independent folding). The simulations of the structures of these two peptides were only partly successful. The most successful folding simulation was that of a 22-residue peptide corresponding to the membrane binding domain of blood coagulation factor VIII (Membind). Three simulations were performed on this peptide and the lowest energy conformation was found to be the most similar to the experimental structure. The conformation of this peptide was determined with a C alpha rms deviation of 4.4 A. Although these simulations were partly successful there are still many unresolved problems, which we expect to be able to address in the next structure prediction experiment.

Algorithms↗
PubMed1994

Cavity mutants of Savinase. Crystal structures and differential scanning calorimetry experiments give hints of the function of the buried water molecules in subtilisins.

The subtilisin molecule possesses several internal water molecules, which may be characterised as an integral part of the protein structure. We have introduced specific mutations (T71I, T71S, T71V, T71A and T71G) at position 71 in the subtilisin variant Savinase from Bacillus lentus. This position is involved in a hydrogen bonded network with several internal water molecules, forming a water channel. The water channel and most of the other internal water molecules are positioned in the interface between two half-domains of the subtilisin molecule. The data presented here indicate that the internal water molecules are structural, and may be the result of trapping during the folding process.

Amino Acids↗
PubMed1994

Humanization of murine monoclonal antibodies through variable domain resurfacing.

Two murine monoclonal antibodies, N901 (anti-CD56) and anti-B4 (anti-CD19), were humanized by a process we call "resurfacing." A systematic analysis of known antibody structures has been used to determine the relative solvent accessibility distributions of amino acid residues in murine and human antibody variable (Fv) regions and has shown that the sequence alignment positions of surface amino acids for human and murine variable region heavy (VH) and light (VL) chains are conserved with 98% fidelity across species. While the amino acid usage at these surface positions creates surface residue patterns that are conserved within species, there are no identical patterns across species. However, surprisingly few amino acid changes need to be made to convert a murine Fv surface pattern to that characteristic of a human surface. Resurfacing was used to change the patterns of surface accessible residues in the Fv regions of the N901 and anti-B4 antibodies to resemble those found on the Fv regions of human antibody sequences. Two different procedures for selecting a human sequence were compared. For anti-B4, a data base of clonally derived human VL-VH sequence pairs was used, while for N901, sequences for VL and VH were independently selected from the Kabat et al. data base [Kabat, E. A., Wu, T. T., Reid-Miller, M., Perry, H. M. & Gottesman, K. S. (1991) Sequences of Proteins of Immunological Interest (DHHS, Washington, DC), 5th Ed.]. Resurfaced N901 and anti-B4 antibodies had apparent affinities for their cell surface ligands that were identical to those of their respective parent murine antibodies. These data provide evidence that, despite the differences in the surfaces of mouse and human Fv regions, it is possible to substitute one for the other while retaining full antigen binding affinity.

Amino Acid Sequence↗
PubMed1994

Comparison of surface accessible residues in human and murine immunoglobulin Fv domains. Implication for humanization of murine antibodies.

Statistical analysis of a database of unique human and murine immunoglobulin heavy chain and light chain variable regions reveals that the precise patterns of exposed residues are different in human and murine antibodies, while most individual surface positions have strong preferences for a small number of residue types. Consideration of these surface patterns alone generates almost identical family groupings for light and heavy chain variable domain sequences to those produced by methods such as those of Kabat et al., where N-terminal framework sequences only are compared, or Tomlinson et al., in which entire variable region nucleotide sequences are used. This unexpected result suggests that the surfaces of V-regions are at least as well conserved as the core framework sequences. Furthermore, using these patterns of human and murine surface residues a novel method for the "humanization" of murine antibodies has been developed and tested.

Algorithms↗
PubMed1994

Antibody design: beyond the natural limits.

Dissection of antibody-antigen interactions requires a knowledge of antibody structure, the ability to model accurately the conformation of antibody-combining sites, and an understanding of the energetic factors governing the interactions. When this understanding has reached the point where the molecular shape and chemical character of a combining site necessary to define a particular specificity and binding requirement can be designed, the antibody repertoire will have been extended 'beyond the natural limits'.

Animals↗
PubMed1994

Mutational and structural analysis of the lectin activity in binding domain 2 of ricin B chain.

The study of the lectin binding sites of ricin B chain and of other homologous members of the small gene family that make up ricin-like molecules has revealed a number of key contact residues involved in sugar binding. In particular, on the basis of data generated by the X-ray crystallographic structure of ricin, comparisons of sequence homologies to other ricin-like molecules and substrate binding studies with these molecules, it has been proposed that His248 of Ricinus communis agglutinin (RCA) B chain may interfere with galactose binding in the second binding domain of that lectin. To test that hypothesis, single binding domain 2 (SBD2) of ricin B chain was expressed as a gene 3 fusion protein on the surface of fd phage to measure directly the effect of mutational changes on this binding site. Replacement of tyrosine with histidine at amino acid position 248 of SBD2 of ricin B chain was shown to reduce lectin activity. The sequences of RCA and ricin B chains were aligned and compared with the tertiary structure of ricin B chain to select various mutations that were introduced as controls in the study. One of these controls, Leu247 to Val247, displayed increased affinity for galactosides. The role of sequence changes is discussed in relation to the structural and functional divergence in these molecules.

Amino Acid Sequence↗
PubMed1992

BIOSITE: a program for the interactive comparison of aligned homologous protein sequences.

A program, BIOSITE, providing for the interactive visual comparison of aligned homologous amino-acid sequences is presented, including an example of its application. The program allows for two types of comparison sequence to be generated: an 'identity' sequence and a 'difference' sequence. These may be used on subsets of sequences and in further comparisons to identify candidate sites involved in a distinct functional property. The program should prove a useful tool for biologists engaged in understanding sequence--function relationships.

Amino Acid Sequence↗
PubMed1990

[Smoking habits among Danish physicians, nurses and midwives in 1989].

Health staff play a central role in distribution of knowledge about the injurious effects of tobacco by influencing the smoking habits of the population. The danish council on smoking and health has therefore chosen health staff as one of the first targets for its activities. One of these was conduct of a questionnaire investigation among a representative section of Danish doctors, nurses and midwives about smoking habits and attitudes to the tobacco problem. The random sample consisted of 2,997 persons, of whom 2,606 (87%) replied to the question about daily smoking habits. In all of the age groups, the frequency of smoking was considerably lower among health staff than in the population as a whole, primarily because many had stopped smoking. The frequency among men was 23% for doctors as compared with 50% in the normal population (age 20-69 years) and 15% for female doctors, 29-30% for nurses and midwives as compared with 46% in the normal population. The frequency of smoking among doctors has decreased considerably since 1980. Very few heavy smokers were found among health staff, particularly among doctors, and many male doctors smoked pipes (47% of the smokers). Even though health staff smoke less than the rest of the population, attention must still be focussed on this group on account of its function for establishing opinions in the health sector.

Adult↗
PubMed1990

Octapeptide segments from the amino terminus of glycophorin A contain the antigenic determinants of the M and N blood groups systems.

Human red blood cells with phenotype N/N and M/M were tested in an agglutination assay with anti-N and anti-M antibodies, respectively. After incubation of the synthesized octapeptide (Leu-Ser-Thr-Thr-Glu-Val-Ala-Met) from the N-amino terminus of glycophorin A, with anti-N antibody, there was significant inhibition of the agglutination of the N-positive cells. There was also inhibition of the agglutination of the M-positive cells with anti-M antibody by the synthesized octapeptide (Ser-Ser-Thr-Thr-Gly-Val-Ala-Met) from the M-amino terminus of glycophorin A. There was no inhibition, however, of the agglutination of M-positive cells with anti-M antibodies by the N-amino terminal octapeptide. Likewise, the M-amino terminal octapeptide did not inhibit agglutination of N-positive cells with anti-N. Because the synthesized octapeptides contained no carbohydrate, the anti-N and anti-M specificity appears to be determined principally by the peptides themselves. Further studies with the use of chimeric peptides indicate that the amino terminal amino acid leucine of N-glycophorin A is a primary determinant of the N antigen, whereas the amino terminal serine of M-glycophorin A is a primary determinant for the M antigen.

Amino Acid Sequence↗
PubMed1989

Colo-pleural fistula.

A case of non-traumatic colo-pleural fistula is recorded for the first time as the cause of a long-standing pleural empyema. The patient was treated with drainage, antibiotics and parenteral nutrition. The fistula was the result of a diverticulitis coli and/or a pancreatitis.

Colonic Diseases↗