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K Harlos

Publications and source records attributed to K Harlos.

53 records · Page 3Linked to original sources
PubMed1990

Crystallization of a soluble form of the rat T-cell surface glycoprotein CD4 complexed with Fab from the W3/25 monoclonal antibody.

The structure of the T-lymphocyte cell surface glycoprotein CD4 is of considerable biological and medical interest. Recombinant rat CD4 expressed in soluble form in mammalian cells and complexed with W3/25 monoclonal Fab fragments formed crystals that diffract to 3.5 A and have the orthorhombic space group P2(1)2(1)2 or P2(1)2(1)2(1). The unit cell has dimensions a = 317 A, b = 161 A and c = 41.8 A and the asymmetric unit consists of two CD4:Fab complexes. These crystals are of suitable quality for X-ray diffraction analysis.

Animals↗
PubMed1987

Structure and order of the protein and carbohydrate domains of prothrombin fragment 1.

The three-dimensional structure of prothrombin fragment 1 has been determined by X-ray crystallography at 3.8 A resolution. The fragment is composed of a number of structural units, some of which are ordered while others are disordered. The ordered part of the structure includes a compact kringle unit, a helical domain and a carbohydrate chain. The kringle structure is organized around a close pair of buried disulfide bridges. One of its carbohydrate chains, that attached to Asn 101, is fully ordered, but the carbohydrate chain attached to Asn 77 appears to be disordered. The calcium binding unit is composed of a disordered part containing all ten gamma-carboxyglutamic acid residues and an ordered part forming the helical domain. The highly conserved residues Phe 41, Trp 42 and Tyr 45, which form a hydrophobic cluster on the first helix, interact around a crystallographic two-fold axis with the equivalent residues in another molecule to form a dimer in the crystal.

Amino Acid Sequence↗
PubMed1987

Conformation and packing properties of membrane lipids: the crystal structure of sodium dimyristoylphosphatidylglycerol.

The conformation and molecular packing of sodium 1,2-dimyristoyl-sn-glycero-phospho-rac-glycerol (DMPG) have been determined by single crystal analysis (R = 0.098). The lipid crystallizes in the monoclinic spacegroup P2(1) with the unit cell dimensions a = 10.4, b = 8.5, c = 45.5 A and beta = 95.2 degrees. There are two independent molecules (A and B) in the asymmetric unit which with respect to configuration and conformation of their glycerol headgroup are mirror images. The molecules pack tail to tail in a bilayer structure. The phosphoglycerol headgroups have a layer-parallel orientation giving the molecules an L-shape. At the bilayer surface the (-) phosphoglycerol groups are arranged in rows which are separated by rows of (+) sodium ions. Laterally the polar groups interact by an extensive network of hydrogen, ionic and coordination bonds. The packing cross-section per molecule is 44.0 A2. The hydrocarbon chains are tilted (29 degrees) and have opposite inclination in the two bilayer halves. In the chain matrix the chain planes are arranged according to a so far unknown hybride packing mode which combines the features of T parallel and O perpendicular subcells. The two fatty acid substituted glycerol oxygens have mutually a - synclinal rather than the more common + synclinal conformation. The conformation of the diacylglycerol part of molecule A and B is distinguished by an axial displacement of the two hydrocarbon chains by four methylene units. This results in a reorientation of the glycerol back bone and a change in the conformation and stacking of the hydrocarbon chains. In molecule A the beta-chain is straight and the gamma-chain is bent while in molecule B the chain conformation is reversed.

Hydrogen Bonding↗
PubMed1987

Deriving the generic structure of the fibronectin type II domain from the prothrombin Kringle 1 crystal structure.

The proposed homology between the fibronectin type II domain and the Kringle domains of blood clotting and fibrinolytic proteins has been examined in three dimensions by substituting the type II sequence into the bovine prothrombin Kringle 1 tertiary structure, determined by X-ray crystallographical methods at 3.8 A. Structural substitution of aligned amino acids of the type II domains and the Kringle produces a compact chain fold and deletions and insertions in the type II sequence are accommodated within the modelled structure. This confirms the structural homology between the two domains and verifies the sequence alignment and common evolution of the type II and Kringle units. The two structures contain homologous hydrophobic cores, centered around the two disulphide bridges which link conserved beta-type strands. Gross differences between the two domains occur in exterior loops and potential functional sites in these regions of the type II structures as found in fibronectin, Factor XII and seminal fluid protein PDC-109 are proposed. We suggest that the domains evolved from a common ancestral protein comprising the hydrophobic core and disulphide arrangement which later diverged to bind different macromolecules through adaptation of the external loops.

Amino Acid Sequence↗
PubMed1983

Metastability and polymorphism in the gel and fluid bilayer phases of dilauroylphosphatidylethanolamine. Two crystalline forms in excess water.

The phase behavior of 1,2-dilauroyl-sn-glycero-3-phosphoethanolamine (DLPE) in excess water has been studied by differential scanning calorimetry and x-ray diffraction. In addition to the usual gel phase L beta of lamellar periodicity 5.07 nm and the fluid bilayer phase L alpha of periodicity 4.5 nm, two distinct crystalline forms may also be spontaneously adopted. The L beta phase is only produced by cooling from L alpha and is metastable, relaxing to one of the crystalline forms on incubation. One crystalline polymorph, designated beta 2, has a lamellar periodicity of 4.55 nm and corresponds to the structure of DLPE crystallized from a variety of organic solvents. The other crystalline polymorph, designated beta 1, has a lamellar periodicity of 3.78 nm, which implies that in this form the hydrocarbon chains are tilted at approximately 40 degrees to the bilayer normal. The beta 2 polymorph is obtained on dispersing crystalline DLPE directly in water at T less than 43 degrees C, on incubation in the L alpha phase at 30 degrees C less than T less than 43 degrees C, or on heating the beta 1 form slowly to T greater than 35 degrees C. The beta 1 polymorph is obtained on incubating the L beta phase at T less than 30 degrees C. By calorimetry, the L beta phase undergoes an endothermic transition (delta H = 15.5 kJ X mol-1 (3.7 kcal X mol-1)) at 30.6 degrees C to the fluid bilayer phase L alpha. The beta 1 phase undergoes an endothermic transition (delta H congruent to 50 kJ X mol-1 (12 kcal X mol-1)) to L alpha at 35 degrees C. The beta 2 phase undergoes an endothermic transition (delta H = 57 kJ X mol-1 (13.7 kcal X mol-1)) to L alpha at 43 degrees C.

Drug Stability↗
PubMed1981

Charge-induced tilt in ordered-phase phosphatidylglycerol bilayers evidence from X-ray diffraction.

X-ray diffraction studies have been performed, as a function of water content, on dipalmitoyl phosphatidylglycerol bilayers, both in the charged state at pH 8.0 and in the protonated state at pH 1.5, using buffers of 1.5 M salt concentration. Measurements were made at 20 degrees C, and the high-angle reflections indicated that the bilayers were in the ordered phase at both pH values. Lamellar diffractions were observed under all conditions studied. THe lamellar repeat reached a limiting value of 62.4 A (6.24 nm) at a water/lipid ratio of 0.24 at pH 8.0, and a limiting value of 67.3 A (6.73 nm) at a water/lipid ratio of 0.22 at pH 1.5. The area per lipid molecule in the plane of the bilayer, deduced from the bilayer thickness and the lipid partial specific volume, is 48 A2 (0.48 nm2) at pH 8.0 and 37 A2 (0.37 nm2) at pH 1.5. The area per molecule in the plane perpendicular to the chain axes, deduced from the X-ray short spacings, is 40.5 A2 (0.405 nm2) at pH 8.0 and 39.2 A2 (0.392 nm2) at pH 1.5. Thus the lipid molecules are tilted by approx. 30 degrees relative to the bilayer normal at pH 8.0, but are essentially untilted at pH 1.5.

Hydrogen-Ion Concentration↗
PubMed1981

Hexagonal phases in phospholipids with saturated chains: phosphatidylethanolamines and phosphatidic acids.

The structure of phospholipids with saturated chains was investigated by differential scanning calorimetry and by X-ray diffraction. It is shown that both phosphatidylethanolamines and phosphatidic acids can exhibit a hexagonal phase at high temperature. The temperature of the transition to the hexagonal phase is dependent on chain length and sodium salt concentration. Increasing the chain length or the sodium salt concentration results in a decrease in the transition temperature. In addition to this transition at high temperature, a calorimetric transition at low temperature, a calorimetric transition at low temperature is detected in some phospholipids.

Calorimetry, Differential Scanning↗
PubMed1980

Influence of calcium on phosphatidylethanolamine. An investigation of the structure at high pH.

The influence of calcium on the structure of 1,2-dihexadecyl-sn-glycero-3-phosphoethanolamine was investiged at pH 8 and 12 by differential scanning calorimetry and by X-ray diffraction. At pH 12, where the amino group of phosphatidylethanolamine is partly deprotonated, two separate lamellar phases are observed at 60 degrees C in the presence of CaCl2; one phase is metastable and the other is present in a crystalline structure. At high temperature (90 degrees C), the small-angle diffraction lines indicate the existence of a hexagonal phase at pH 8. At pH 12 a hexagonal phase is detected only in the presence of calcium.

Calcium↗
PubMed1980

Influence of calcium on phosphatidylglycerol. Two separate lamellar structures.

The influence of calcium on the structure of rac-1,2-ditetradecylglycerol-3-phosphoglycerol is investigated by differential scanning calorimetry and by X-ray diffraction. It is shown that 1 M CaCl2 (pH 4.6) induces two separate lamellar phases in the same sample at 20 degrees C. These two phases can be clearly distinguished by their X-ray diffraction patterns. The type of phase observed depends on the pretreatment of the sample. At high temperature (90 degrees C), when the hydrocarbon chains are in the disordered state, the small angle reflections are in the ratio 1:1/square root 3:1/2 and thus indicate the presence of a hexagonal phase.

Calcium↗
PubMed1980

Charge-induced pretransition in phosphatidylethanolamine multilayers. The occurrence of ripple structures.

The influence of pH and ionic strength on the phase transition behavior of 1,2-dihexadecylphosphatidylethanolamine was studied calorimetrically. In the range of ionic strength from 0.75 to 1.5 M NaCl at pH larger than 13, where the amino group of the phosphatidylethanolamine is in the deprotonated state, resulting in one negative charge per lipid molecule, the calorimetric scan shows a pretransition before the main transition. Accompanying freeze-fracture electron microscopic studies on these preparations in the temperature range between the pre- and main transitions show a regular suface, the so-called ripple structure. These are comparable with the structures seen in phosphatidylcholine-water systems at temperatures between the pre- and main transition.

Calorimetry↗
PubMed1979

Influence of pH on phosphatidic acid multilayers. A rippled structure at high pH values.

The influence of pH on the structure of 1,2-(ditetradecyl)-phosphatidic acid was investigated by differential scanning calorimetry and freeze-fracture electron microscopy. At pH 13.5--14 (2.6 M K+), where phosphatidic acid has two negative charges, calorimetric scans show a small transition (pretransition) below the main phase transition temperature. Freeze-fracture studies of the same dispersions reveal regular band patterns (so-called ripples) in the plane of the bilayers, when the lipid is quenched from below the main phase transition temperature. This rippled structure is similar to the well-known rippled structure of phosphatidylcholines.

Calorimetry, Differential Scanning↗
PubMed1979

Electrostatic interactions at charged lipid membranes. Electrostatically induced tilt.

The changes in bilayer structure induced by surface charges in the case of an ionizable lipid were studied by X-ray diffraction, Raman spectroscopy, and film-balance measurements. With increasing surface charge in the ordered phase, the X-ray results show a decrease in bilayer thickness, whereas the hydrocarbon chain packing stays essentially constant, the Raman data signify that the internal chain ordering does not change, and the monolayer studies show a lateral expansion of the bilayer. These results are interpreted in terms of a tilt of the chains caused by the surface charges on the polar heads. The tilt angle between the direction of the chains and the bilayer normal is obtained by a detailed theoretical evaluation. The tilt allows for a better understanding of the electrostatically induced shift of the phase transition temperature and of the shift induced by the binding of water in the case of lecithin in contrast ethanolamine.

Hydrogen-Ion Concentration↗
PubMed1978

Pretransitions in the hydrocarbon chains of phosphatidylethanolamines. A wide angle X-ray diffraction study.

The hydrocarbon chain packing of fully hydrated phosphatidylethanolamine multilayers is investigated by X-ray diffraction. An analysis of the wide angle reflections (short spacings) as a function of temperature indicates that, apart from the well-known ordered-disordered lipid phase transition, a second transition takes place at lower temperatures. This transition, which is in the present paper referred to as the pretransition, is characterized by a transformation of the hydrocarbon chain packing. A first model for the chain lattice is presented, which gives rise to the expectation that similar pretransitions might be found with other phospholipids.

Chemical Phenomena↗
PubMedDate not supplied

Vitamin K-dependent blood coagulation proteins form hetero-dimers.

The later stages of the blood coagulation cascade are characterized by the presence of vitamin K-dependent proteins and their involvement in membrane-bound, multi-protein converting complexes with an essential requirement for calcium ions. Specific interactions between zymogens and activating enzymes have not yet been identified. Here we describe a crystallographic study of prothrombin fragment 1 (residues 1-156 of prothrombin) which indicates that vitamin K-dependent coagulation proteins have specific association sites that allow them to form hetero-dimers. The calcium-induced formation of a hetero-dimer between fragment 1 and factor X is demonstrated by cross-linking. Such hetero-dimers of vitamin K-dependent proteins could be significant in the coagulation system.

Blood Coagulation↗