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K Chandrasekhar

Publications and source records attributed to K Chandrasekhar.

At least 19 recordsLinked to original sources
PubMed1991

Solution conformational preferences of immunogenic peptides derived from the principal neutralizing determinant of the HIV-1 envelope glycoprotein gp120.

With standard one- and two-dimensional proton NMR techniques, a common structural motif has been identified in water solutions of short peptide sequences derived from the envelope glycoprotein gp120 of HIV-1. Three peptides of lengths 12, 24, and 40 residues (termed RP342, RP142, and RP70, respectively) were synthesized, each containing a central amino acid sequence common to many HIV-1 isolates. In addition, RP70 contained a disulfide bond between cysteine residues close to the ends of the molecule, forming a loop that is thought to constitute an important structural and immunological component of the intact glycoprotein. Peptides RP70 and RP142 showed evidence for the presence of a significant population of conformations containing a beta-turn in the conserved sequence Gly-Pro-Gly-Arg. Strong nuclear Overhauser effect (NOE) connectivities were observed between the amide protons of the arginine and the adjacent glycine. A weak NOE connectivity was observed between the C alpha H of the proline residue and the NH of the Arg [a d alpha N(i,i + 2) NOE connectivity], confirming the presence of a conformational preference for a turn conformation in this sequence. The remainder of the peptide showed evidence of conformational averaging: no NMR evidence for a uniquely folded structure was obtained for any of the peptides in water solution. Circular dichroism (CD) spectra indicated that no ordered helix was present in water solutions of RP70, although a CD spectrum that indicated the presence of approximately 30% helix could be induced by the addition of trifluoroethanol.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1991

Two-dimensional proton-NMR studies on a hybrid peptide between cecropin A and melittin. Resonance assignments and secondary structure.

A hybrid peptide of cecropin A and melittin was investigated by two-dimensional 1H-NMR at pH 5.8 in aqueous solution with 30% (by vol.) hexafluoroisopropanol. The peptide contains 26 amino acids, is a combination of the first 13 residues of each of the two parent peptides, CA(1-13)M(1-13) identical to CAM(1-26) and has an amidated C terminal. This peptide was recently synthesized [Boman, H.G., Wade, D., Boman, I.A., Wåhlin, B. & Merrifield, R.B. (1989) FEBS Lett. 259, 103-106] and shown to have strong antibacterial activity but to be harmless towards erythrocytes. All resonances of the main chain and side chain beta-protons are assigned except for those of the N-terminal lysine. Several medium range NOE connectivities were observed showing two separated alpha-helices, involving residues 4-12 and 16-26. The JNH alpha-coupling constants in these sections support the conclusion. From the exchange rates of the NH protons it is concluded that the alpha-helix of residues 16-26 is much more stable than the other helix. The circular dichroism data indicates about 30% less alpha-helix character than the NMR data. A reduced contribution to the ellipticity from the unstable helix is suggested. The chemical-shift differences between the two parts of the hybrid and the respective parent peptides are larger for the cecropin part than for the melittin part. For the latter, residues 17-26 of the hybrid are proposed to have a secondary structure very similar to that of residues 4-13 of melittin.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1991

Assignment of the 15N NMR spectra of reduced and oxidized Escherichia coli thioredoxin.

As a necessary first step in the use of heteronuclear correlated spectra to obtain high resolution solution structures of the protein, assignment of the 15N NMR spectra of reduced and oxidized Escherichia coli thioredoxin (Mr 12,000) uniformly labeled with 15N has been performed. The 15N chemical shifts of backbone amide nitrogen atoms have been determined for both oxidation states of thioredoxin using 15N-1H correlated and two-dimensional heteronuclear single-quantum coherence (HSQC) TOCSY and NOESY spectra. The backbone assignments are complete, except for the proline imide nitrogen resonances and include Gly33, whose amide proton resonance is difficult to observe in homonuclear 1H spectra. The differences in the 15N chemical shift between oxidized and reduced thioredoxin, which occur mainly in the vicinity of the two active site cysteines, including residues distant in the amino acid sequence which form a hydrophobic surface close to the active site, are consistent with the differences observed for proton chemical shifts in earlier work on thioredoxin.

Amino Acid Sequence↗
PubMed1990

Direct methods with single isomorphous replacement data. I. Reduction of systematic errors.

The direct-methods procedure for single isomorphous replacement (SIR) data [Hauptman (1982). Acta Cryst. A38, 289-294], as modified by Fortier, Moore & Fraser [Acta Cryst. (1985), A41, 571-577] has been implemented and tested with a large number of known structures. It was found that the modified procedure greatly reduces the bias toward 'unresolved' SIR invariant values associated with estimates of 0 or pi, but does not remove it entirely. If the heavy atoms are not in a centrosymmetric array the centroid of the distribution of invariant estimates is not centered on true protein values, but is biased toward conventional SIR values by up to 15 degrees, thus errors in the estimates are not random but systematic. When the heavy atoms are in a centrosymmetric array (or single heavy-atom site in space group P21), the distribution of estimates is often sharply bimodal, with peaks centered at both true invariant values and pure 'unresolved' SIR values. Simple procedures are given which can be applied in both situations to reduce significantly the bias with no overall loss of accuracy. An additional correction factor is then described which can be used to remove nearly all of the bias, and improve the accuracy as well. The result is that errors in the corrected invariant estimates are small in magnitude, but are now also random instead of systematic. Since the number of estimates greatly exceeds the number of phases, the remaining random errors should have little impact in phasing processes.

Bence Jones Protein↗
PubMed1985

Preliminary crystallographic study of an L-asparaginase from Vibrio succinogenes.

Crystals of an L-asparaginase from Vibrio succinogenes were obtained with the hanging drop method from ammonium sulphate-containing solutions. The crystals belong to the orthorhombic space group P22(1)2(1) with unit cell dimensions of a = 71.3 A, b = 85.8 A, c = 114.0 A, and contain two tetrameric enzyme molecules per unit cell. There are two subunits in the asymmetric unit; a molecular dyad is coincident with the crystallographic dyad. The crystal lattice is similar to that reported for an Escherichia coli asparaginase. Rotation function calculations have revealed that the V. succinogenes enzyme has 222 point group symmetry in the crystal. The second and third molecular dyads differ, however, from the corresponding E. coli asparaginase dyads by approximately 40 degrees. The crystals diffract to at least 2.2 A resolution and are suitable for X-ray crystallographic structure determination.

Asparaginase↗
PubMed1980

Artificial spawning effected in the fresh water teleost, Cyprinus carpio by clomiphene citrate.

Triweekly i.m. injections of clomiphene citrate (group 1, 25 micrograms/0.5 ml and group II, 50 micrograms/1.0 ml) were administered for a period of 3 months during the preparatory period to female fresh water teleosts exhibiting ovarian recrudescence, while a control group received 0.5 ml of physiological saline throughout the period of experimentation i.e., from February through April. 50 micrograms clomiphene citrate treatment brought about a steady increase in ovarian size, and oocytes began to enlarge and mature and finally ovulation took place in April. This is 4 months ahead of their normal occurrence.

Animals↗
PubMed1979

Light and electron microscopical investigations on the tanycyte differentiation during the perinatal period in the rat.

The differentiation of tanycytes was studied light and electron microscopically during the perinatal period in rats, the time when functional connections between hypothalamus and hypophysis are established. The 3rd ventricle is slit-like between 16 and 18 days of the prenatal period. Its wall is formed by intensively proliferating matrix cells with apical processes, ovoid perikarya and a basal process. The ventral region of the 3rd ventricle becomes funnel-shaped on the 20th day of the prenatal period. As the cells differentiate, the apical process becomes shorter and broader. Moreover, on day 20 of prenatal life cells without apical processes appear. Their number increases during the postnatal period. The concentration of endoplasmic reticulum, mitochondria, polysomes, lipid droplets, dense bodies (lysosomes), lamellated and multivesicular bodies increases. Initially the cells are similar but from the 3rd day of postnatal life differentiation occurs in different regions of the infundibular recess. After the 5th day, there are no marked changes in the structure and distribution of these cells.

Animals↗
PubMed1979

Quantitative protein changes in the hypothalamic neurons of pubertal male rats, castrated neonatally.

The effect of neonatal castration of male rats on the sexual differentiation of the hypothalamus at puberty was studied. Male rats were castrated on days 1, 5 and 7 after birth. Their brains were processed for study on days 83-85. The neurons and cell nuclei of the preoptic area, mediobasal and ventromedial nuclei were assessed for changes in cell and nuclear sizes and dry weight (calculated using interferometric methods). Neonatal castration resulted in size as well as dry weight increase in the neurons of the anterior and mediobasal hypothalamus. The dry weight increased by 34% (P less than 0.001) in the medial preoptic area, by 25% (P less than 0.001) in the arcuate neurons and by 22% (P less than 0.001) in the ventromedial nucleus. The cell nuclei exhibited perceptible weight increase too--in the medial preoptic area 68% (P less than 0.001); 55% in the arcuate neurons (P less than 0.001), and 39% in the ventromedial region. The weight and size increases in neonatally castrated males were equal to those of females of the same age. In rats castrated on day 7, the cell sizes and dry weights of the ventromedial nucleus increased but the cell nuclei exhibited only little change. It is assumed that the changes in the dry weight may be the result of increased synthetic processes in these groups of neurons which are connected with the tonic and cyclic release of gonadotropins. These changes also point to the hypothalamic differentiation shifting to the female type in the absence of the inducing effect of androgens.

Animals↗