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D Tsernoglou

Publications and source records attributed to D Tsernoglou.

14 recordsLinked to original sources
PubMed1992

Refined structure of the pore-forming domain of colicin A at 2.4 A resolution.

The E1 subgroup (E1, A, B, IA, IB, K and N) of anti-bacterial toxins called colicins is known to form voltage-dependent channels in lipid bilayers. The crystal structure of the pore-forming domain of colicin A from Escherichia coli has been refined to the diffraction limit of the crystals at 2.4 A resolution by means of molecular dynamics and restrained least-squares methods to a conventional R-factor of 0.18 for all data between 6.0 and 2.4 A resolution. The polypeptide chain of 204 amino acid residues consists of ten alpha-helices organized in a three-layer structure. The helices range in length from 9 to 23 residues with an average length of 125 residues. The packing arrangement of the helices has been analysed; the packing is different from that observed in four-helix bundle proteins. The sites of 83 water molecules have been located and refined. Analysis of the structure provides insights into the mechanism of formation of a voltage-gated channel by the protein. Although it is proposed that substantial tertiary structural changes occur during membrane insertion, the secondary structural elements remain conserved. This idea has been proposed recently for a number of other protein-membrane events and thus may have more general applicability.

Amino Acid Sequence↗
PubMed1991

Crystallization and preliminary X-ray analysis of phosphoporin from the outer membrane of Escherichia coli.

Phosphoporin is a pore-forming transmembrane protein that spans the outer membrane of Escherichia coli and facilitates the diffusion of phosphates and phosphorylated compounds. Phosphoporin has been crystallized in several different crystal forms, although only one appears to be suitable for X-ray analysis. These crystals, which are hexagonal plates, diffract X-rays to 3 A resolution and belong to the space-group P6(3)22, with unit cell dimensions a = b = 121 A and c = 111 A.

Bacterial Outer Membrane Proteins↗
PubMed1991

A common channel-forming motif in evolutionarily distant porins.

Four new crystal packings of Escherichia coli porins are presented (phosphoporin, maltoporin, and two crystal forms of matrix porin). These were determined by molecular replacement methods using a polyalanine trial model acquired from the refined coordinates of porin from Rhodobacter capsulatus. The successful molecular replacement shows that the dominant motif found in R. capsulatus porin (a 16-stranded antiparallel beta-barrel) also applies to the E. coli porins, despite the lack of significant amino acid sequence homology. A 30 degrees-40 degrees tilt of the beta-strands with respect to the membrane normal was derived from the intensity distributions in the X-ray diffraction patterns for each porin studied, stressing their similarity. In view of the evolutionary distance between enteric and photosynthetic bacteria, the antiparallel beta-barrel may have significance as a basic structural motif for the formation of bacterial membrane channel structures.

Bacterial Outer Membrane Proteins↗
PubMed1990

Colicins: prokaryotic killer-pores.

Colicins are plasmid-encoded protein antibiotics which kill bacteria closely related to the producing strain (generally Escherichia coli). The study of the function of colicins has revealed many features which reflect common targeting and translocation mechanisms with bacteriophages and toxins. Like many toxins, colicins are composed of structural domains specialized in one of the different steps of the activity, targeting, translocation and killing. The major group comprises those colicins which permeabilize the cytoplasmic membrane, thereby destroying the cell's membrane potential. These colicins form well-defined voltage-gated ion channels in artificial membranes. The scope of this review is to describe some of the more recent findings concerning the structure and mode of action of pore-forming colicins with a special attention to models of membrane insertion and pore structure based on the recently determined three-dimensional structure of the pore-forming domain of colicin A.

Amino Acid Sequence↗
PubMed1979

Temperature-dependent X-ray diffraction as a probe of protein structural dynamics.

X-ray diffraction at four temperatures from 220 to 300 K coupled with crystallographic refinement yields the mean-square displacements and conformational potentials of all 1,261 non-hydrogen atoms of metmyoglobin. The results are interpreted to indicate a condensed core around the haem, semi-liquid regions towards the outside and a possible pathway for ligands. It is concluded that X-ray diffraction can provide the spatial distribution of the dynamic features of a protein.

Animals↗
PubMed1977

Protein sequencing by computer graphics.

A computer graphics system has been used to fit a putative sequence to an electron density map of a sea snake neurotoxic protein at 2.2 A resolution. The complete sequence of this small protein could be determined from the map with very little ambiguity. In two places probable errors in the published chemical sequence were detected. This is the first instance in which a complete three-dimensional structure was solved with the use of computer graphics alone, without the construction of a physical model.

Amino Acid Sequence↗
PubMed1977

Three-dimensional structure of neurotoxin a from venom of the Philippines sea snake.

The crystal structure of neurotoxin a from the venom of Philippines sea snake Laticauda semifasciata has been determined at 2.5 A resolution by x-ray diffraction. Comparison with the structure of neurotoxin b from the same source indicates that the two toxins differ only by substitution at His 26. Earlier chemical work had suggested a difference in chain length and considerable differences in amino acid composition. The difference between our a and b toxins is the same as that reported from sequence analysis for the related erabutoxins a and b from Japanese sea snake, and suggests that the Philippines toxin may be identical to erabutoxin. The replacement of His 26 by a shorter side-chain in toxin a has no effect on the structure of the rest of the molecule. In particular, the protruding loop, which we believe interacts with the acetylcholine receptor, is not affected, even though His 26 is in the loop.

Animals↗
PubMed1976

Crystal structure of elastase-substrate complex at -- 55 degrees C.

The structure of a specific acyl-enzyme intermediate in the elastase-catalysed hydrolysis of N-carbobenzoxy-L-alanyl-p-nitrophenol ester has been determined by X-ray diffraction at 3.5 A resolution. The acyl-enzyme was stabilised by cooling the crystal to --55 degrees C during substrate addition and data collection.

Binding Sites↗