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L Sjölin

Publications and source records attributed to L Sjölin.

33 records · Page 2Linked to original sources
PubMed1988

Preliminary crystal structure of Acinetobacter glutaminasificans glutaminase-asparaginase.

The preliminary structure of a glutaminase-asparaginase from Acinetobacter glutaminasificans is reported. The structure was determined at 3.0-A resolution with a combination of phase information from multiple isomorphous replacement at 4-5-A resolution and phase improvement and extension by two density modification techniques. The electron density map was fitted by a polypeptide chain that was initially polyalanine. This was subsequently replaced by a polypeptide with an amino acid sequence in agreement with the sizes and shapes of the side chain electron densities. The crystallographic R factor is 0.300 following restrained least squares refinement with data to 2.9-A resolution. The A. glutaminasificans glutaminase-asparaginase subunit folds into two domains: the aminoterminal domain contains a five-stranded beta sheet surrounded by five alpha helices, while the carboxyl-terminal domain contains three alpha helices and less regular structure. The connectivity is not fully determined at present, due in part to the lack of a complete amino acid sequence. The A. glutaminasificans glutaminase-asparaginase structure has been used successfully to determine the relative orientations of the molecules in crystals of Pseudomonas 7A glutaminase-asparaginase, in crystals of Vibrio succinogenes asparaginase, and in a new crystal form of Escherichia coli asparaginase (space group 1222, one subunit per asymmetric unit).

Acinetobacter↗
PubMed1986

Multiple conformations of amino acid residues in ribonuclease A.

The highly refined 1.26 A structure (R = 0.15) of phosphate-free bovine pancreatic ribonuclease A was modeled with 13 residues having discrete multiple conformations of side chains. These residues are widely distributed over the protein surface, but only one of them, Lys 61, is involved in crystal packing interactions. The discrete conformers have no unusual torsion angles, and their interactions with the solvent and with other atoms of the protein are similar to those residues modeled with a single conformation. For three of the residues--Val 43, Asp 83, and Arg 85--two correlated conformations are found. The observed multiple conformations on the protein surfaces will be of significance in analyzing structure-function relationships and in performing protein engineering.

Amino Acids↗
PubMed1984

Structure of bovine pancreatic trypsin inhibitor. Results of joint neutron and X-ray refinement of crystal form II.

The structure of form II crystals of bovine pancreatic trypsin inhibitor has been investigated by joint refinement of X-ray and neutron data. Crystallographic R factors for the final model were 0.200 for the X-ray data extending to 1 A resolution and 0.197 for the 1.8 A neutron data. This model was strongly restrained, with 0.020 A root-mean-square (r.m.s.) departure of bond lengths from their ideal values and 0.019 A r.m.s. departure of planar groups from planarity. The resulting structure was very similar to that of crystal form I (r.m.s. deviation for main chain atoms was 0.40 A); nevertheless larger deviations were observed in particular regions of the chain. Twenty out of 63 ordered water molecules occupy similar positions (deviation less than 1 A) in both models. Eleven amide hydrogens were found to be protected from exchange after three months of soaking the crystals in deuterated mother liquor at pH 8.2. Their locations were in excellent agreement with the results obtained by two-dimensional nuclear magnetic resonance, but the rates of exchange are much lower in the crystalline state.

Amino Acid Sequence↗
PubMed1983

Crystallization and preliminary X-ray diffraction data of two crystal forms of bovine heart creatine kinase.

Two crystal forms of bovine heart creatine kinase, which are suitable for X-ray diffraction studies, have been grown at room temperature using 2-methyl-2,4-pentanediol as the precipitant at pH 7.2. The space group of the orthorhombic form is P2(1)2(1)2, with unit cell dimensions a = 133 A, b = 128 A and c = 65 A, and there is one dimeric molecule in the asymmetric unit. The space group of the tetragonal form is P4(2)2(1)2, with unit cell dimensions a = b = 132 A and c = 75 A, with one subunit in the asymmetric unit. The tetragonal crystals diffract to at least 2.0 A resolution.

Animals↗
PubMed1983

Structure of ribonuclease A: results of joint neutron and X-ray refinement at 2.0-A resolution.

The structure of ribonuclease A has been refined jointly with the neutron and X-ray data extending to 2.0 A. The results of an earlier X-ray refinement provided the starting model [Wlodawer, A., Bott, R., & Sjölin, L. (1982) J. Biol. Chem. 257, 1325-1332]. The final R factors were 0.159 (X-ray) and 0.183 (neutron) for a model containing all of the atoms expected in the protein, 128 waters, and a phosphate molecule in the active site. The joint refinement necessitated modifications in the orientation of a number of side chains, including the catalytically active lysine-41, which is now thought to form a salt link to the phosphate. Major modifications of the previous model of the bound solvent were necessary. The refinement of all atom occupancies with the neutron data only provided the information about the amide hydrogen exchange. A fourth of all amide hydrogens were found to be at least partially protected from exchange after a year of exchange with D2O.

Amino Acid Sequence↗
PubMed1983

Active site of RNase: neutron diffraction study of a complex with uridine vanadate, a transition-state analog.

A complex of RNase A with a transition-state analog, uridine vanadate, has been studied by a combination of neutron and x-ray diffraction. The vanadium atom occupies the center of a distorted trigonal bipyramid, with the ribose oxygen O2' at the apical position. Contrary to expectations based on the straightforward interpretation of the known in-line mechanism of action of RNase, nitrogen NE2 of histidine-12 was found to form a hydrogen bond to the equatorial oxygen O8, while nitrogen NZ of lysine-41 makes a clear hydrogen bond to the apical oxygen O2'. Nitrogen ND1 of histidine-119 appears to be within a hydrogen-bond distance of the other apical oxygen, O7. Two other hydrogen bonds between the vanadate and the protein are made by nitrogen NE2 of glutamine-11 and by the amide nitrogen of phenylalanine-120. The observed geometry of the complex may necessitate reinterpretation of the mechanism of action of RNase.

Animals↗
PubMed1983

Kinetics of the TTX sensitive Na+ current in the slowly adapting lobster stretch receptor neurone.

The kinetics of the TTX sensitive Na+ current (INa) in the slowly adapting lobster stretch receptor neurone were investigated in sub- and near-threshold voltage regions using electrophysiological and pharmacological techniques. In dynamic conditions INa was found to display both fast and slow reactions. These were attributed to a fast Hodgkin-Huxley type of Na activation and inactivation, and a slow type of Na inactivation, respectively. In stationary conditions the voltage dependence of the slow Na inactivation was shifted in a depolarizing direction by increasing, and in a hyperpolarizing direction by decreasing the extracellular Ca++ concentration. From this finding as well as from its kinetic properties the slow Na inactivation was classified as a genuine gating process. The processes of fast Na activation and inactivation were too fast for a dynamic analysis with the recording technique available. An estimate of their stationary voltage dependence could however be obtained in a voltage range from about -80 to about -50 mV. The experimental findings were used for the formulation of a mathematical description of INa in the present preparation based on constant field and state transition theories.

Adaptation, Physiological↗
PubMed1982

The refined crystal structure of ribonuclease A at 2.0 A resolution.

This paper describes the structure of bovine pancreatic ribonuclease A, refined by a restrained parameter least squares procedure at 2.0 A resolution, and rebuilt using computer graphics. The final agreement factor (formula see text) is 0.159. The positions of the 951 main chain atoms have been determined with an estimated accuracy of 0.17 A. In addition, the model includes a phosphate group in the active site and 176 waters, many of them with partial occupancy. The bond lengths in the refined structure of RNase A differ from the ideal values by an overall root mean square deviation of 0.022 A; the corresponding value for angle distances is 0.06 A. The root mean square deviation of planar atoms from ideality is 0.017 A, and root mean square deviation of the peptide torsion angles from 180 degrees is 3.4 degrees. The model is in good agreement with the final difference Fourier maps. Two active site histidines, His 12 and His 119, form hydrogen bonds to the phosphate ion. His 119 is also hydrogen bonded to the carboxyl of ASp 121 and His 12 to the carbonyl of Thr 45. The structure of the RNase A is very similar to that of RNase S, particularly in the active site region. The root mean square discrepancy of all atoms from residues 1 to 16 and 24 to 123 is 1.06 A and the root mean square discrepancy for the active site region is 0.6 A.

Animals↗
PubMed1982

Hydrogen exchange in RNase A: neutron diffraction study.

Hydrogen exchange has been studied in a single crystal of RNase A [ribonuclease (pancreatic), EC 3.1.27.5] in the course of a neutron structure investigation. Refinement of the occupancies of amide hydrogens provided information about the kind of isotope present in each site and also provided estimates of the errors associated with the measurement. Twenty-eight of the 120 peptide amide hydrogens were found to be at least partially protected from exchange during approximately 1 year required for crystal preparation and data collection. Most of the protected hydrogens were involved in hydrogen bonds with main-chain carbonyl groups. A contiguous region of the beta-sheet containing residues 75, 106--109, 116, and 118 had a large number of protected hydrogens, indicating its low flexibility and the lack of accessibility to solvent. Residues 11--13 from the alpha-helix near the amino terminus were protected, in good agreement with a model of cooperative unwinding of this helix, starting from the free (amino) end.

Animals↗
PubMed1981

Orientation of histidine residues in RNase A: neutron diffraction study.

Difference Fourier maps have been calculated at 2.8-A resolution by using neutron diffraction data obtained from a single crystal of RNase A. The phases were derived from a model resulting from the joint refinement of x-ray and neutron data at 2.0-A and 2.8-A resolution, respectively. The orientation of histidine-48 assumed during the refinement of the x-ray model at 2.5 A was confirmed, whereas the other three histidines had to be rotated around C beta--C gamma bonds in order to agree with the neutron difference Fourier maps. In the final model, histidine-12 is clearly hydrogen bonded to the carbonyl oxygen of threonine-45 and to the oxygen of the inorganic phosphate, and histidine-119 is bonded to another oxygen of the phosphate and to the oxygen OD1 of aspartic acid-121.

Crystallization↗
PubMed1981

Subthreshold and near-threshold membrane currents in lobster stretch receptor neurones.

1. The ion currents in the slowly and rapidly adapting stretch receptor neurone of lobster were investigated with respect to their nature and stationary kinetics in sub- and near-threshold voltage regions using electrophysiological and pharmacological techniques. 2. In both neurones the following currents were identified: (a) a tetrodotoxin-sensitive Na current, (b) a tetraethylammonium and 4-aminopyridine-sensitive K current, (c) a Co (or Mn)-sensitive Ca-dependent K current, (d) an ouabain-sensitive pump current and (e) a remaining leak current carried mainly by Na, K and Cl. 3. In suprathreshold voltage regions the balance between the individual membrane currents leads to the formation of a stationary negative conductance (negative slope in the voltage dependence of the ionic current) in the slowly, but not in the rapidly adapting cell. 4. These observations are compatible with the fact that during prolonged suprathreshold stimulation a stationary low frequency impulse during is possible in the slowly adapting cell, whereas in the rapidly adapting cell it is not.

Aminopyridines↗