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Biomedical subjects

H Klump

Publications and source records attributed to H Klump.

At least 37 records · Page 2Linked to original sources

Small cytoplasmic RNAs from rat liver mRNP particles. Studies on their structure and function.

Free cytoplasmic 40S mRNP particles from rat liver were treated with EDTA and separated into two populations of RNP particles with sedimentation maxima of 20S and 35S, respectively. A characteristic set of distinct scRNAs is found for 20S and 35S RNP particles. The sequences of two of the most abundant scRNAs from 20S RNP particles with chain lengths of 104 (alpha 1-RNA) and 124 (beta 1-RNA) nucleotides, respectively, are presented. alpha 1-RNA shows a high sequence homology to the 3'-end of 18S rRNA. Since alpha 1-RNA carries a cap, it cannot be a degradation product of 18S rRNA. The beta 1-RNA is strongly post-transcriptionally modified, but uncapped. When the individual scRNAs of 20S and 35S RNP particles isolated from preparative polyacrylamide gels were assayed for their capability to inhibit in vitro protein synthesis, several potent translational inhibitory RNAs were detected. Particularly, the scRNAs of 147,203 and 263 nucleotide length associated with the 35S RNP particles turned out to be strong inhibitors of protein synthesis.

Animals↗

Structural and thermodynamic investigation of the Chilo iridescent virus (Iridovirus type 6).

Structural features and thermodynamic parameters of the complete Chilo iridescent virus (Iridovirus type 6) and its constituents, isolated from the larvae of Galleria mellonella, were evaluated by means of UV spectroscopy and microcalorimetry. It can be demonstrated that the viral DNA is attached to the coat protein in a chromatin-like fashion, which is preserved after disruption of the virus by low temperature or partial digestion of the coat protein with proteinase K. At elevated temperature however the viral DNA is denaturated irreversibly. The coat protein appears to remain in its native state during the DNA transition and its own thermal denaturation profile shows its independence from the DNA denaturation.

DNA, Viral↗

Demonstration of G . U wobble base pairs by Raman and IR spectroscopy.

When guanine and uracil form hydrogen bonds in the pairing scheme first proposed by Crick one would expect that poly(A,G) will form an unperturbed double helix with poly U at room temperature in a dilute electrolyte solution (0.1 M NaCl). We have demonstrated by Raman- and IR-spectroscopy that the secondary structure of poly(A.G) . poly U is very similar to the structure of poly A . poly U; only the thermal stability of the double helix seems slightly lower than the stability of poly A . poly U, whereas the average helix length is unaffected by the dispersed G . U base pairs. From our input ratio of guanine and adenine we estimate that about every fourth base pair is a wobble pair.

Journal Article↗

Kinetics of the helix-coil transition of a polypeptide with non-ionic side groups, derived from ultrasonic relaxation measurements.

Ultrasonic absorption and velocity dispersion curves have been measured in the temperature induced helix-coil transition range of poly-N5-(3-hydroxypropyl)-L-glutamine in a methanol/water mixture. The results clearly reflect an effect due to the kinetics of the conformational conversion. A practically single relaxation time is observed which passes through a maximum when plotted versus the degree of transition. This maximum occurs at definitely less than 50% helix as predicted for by the theory for the comparatively short chain length involved here. The results are discussed in relation to previous theoretical and experimental findings.

Calorimetry↗

Thermal denaturation of nucleosomal core particles.

Thermal denaturation of very homogeneous preparations of core particles from chicken erythrocyte chromatin is studied by several techniques. The change in absorbance, which is very closely paralleled by changes in heat capacity, which is very closely paralleled by changes in heat capacity, is a biphasic process with inflexions at 60 degrees C and 74 degrees C. In contrast, isolated DNA of the same length denatures in a single transition around 44 degrees C. Monitoring the circular dichroism of the cores during thermal denaturation reveals biphasic changes in the secondary structure of the DNA, preceding the base unstacking by 10 degrees C in the first and 3 degrees C in the second phase. However, measurable alterations in the secondary structure of the histones are confined to the second phase with a melting temperature at 71 degrees C. Increase in the ionic strength of the buffer from 1 mM to 10 mM leads to almost monophasic melting curves as measured by absorbance and CD, while not causing any measurable conformational changes at room temperature. The melting of core particles is interpreted as a denaturation of about 40 base pairs in the first phase, followed by a massive breakdown of the native structure of a tight histone-DNA complex, which frees the remaining 100 base pairs for unstacking.

Animals↗

Calorimetric studies on viroids.

Thermodynamic studies on highly purified viroid preparations were carried out with the help of a very sensitive adiabatic microcalorimeter. Parallel to the change of UV-absorption at 260 nm as a function of temperature, the additional heat capacity of the dilute viroid solution rises sharply within the melting interval, reaches a maximum at T = Tm and declines to a baseline again when the temperature is increased further. From the peak area the molar transition enthalpy can be calculated. The transition enthalpies of citrus exocortis viroid and cucumber pale fruit viroid are 4200 kJ/mol and 3930 kJ/mol, respectively. The calorimetric results are compared to the results obtained from melting studies using UV-absorption.

Calorimetry, Differential Scanning↗

Calorimetric measurements of the transition enthalpy of DNA in aqueous urea solutions.

The helix-coil equilibrium of DNA is delicately affected by the nature of the solvent. In this investigation the helical secondary structure was destabilized by an increasing concentration of urea. We found a linear dependence of the transition enthalpy deltaH on the urea concentration for calf thymus DNA as well as for salmon sperm DNA.

Animals↗

Thermodynamic values of the helix-coil transition of DNA in the presence of quaternary ammonium salt.

The knowledge of the enthalpy and entropy of the helix-coil transition in DNA is necessary for the understanding of the stabilization of its native conformation in solution. Reported here is the transition temperature Tm, the transition enthalpy deltaH, determined with the help of an adiabatic scanning calorimeter, the transition entropy deltaS and the breadth of the helix-coil transition as a function of tetramethyl and tetraethyl ammonium chloride concentration.

Animals↗

A calorimetric study of polyguanylic acid at neutral pH.

The structure of polyguanylic acid (poly G) at neutral pH has been studied by optical and calorimetrical methods. It can be shown that diverging from earlier findings Poly G reversibly undergoes a cooperative thermal transition. Thermal denaturation curves are recorded at 253 nm as a function of the sodium ion concentration. The denaturation enthalpy of poly G in dilute aqueous solution is determined to 2.2 kcal/mole g. It is concluded, that the part of the ordered poly G structure, which gives rise to a temperature dependent cooperative transition, arises from stacking interactions of adjacent bases in the single strand.

Calorimetry↗

Calorimetric studies of the interaction between DNA and poly-L-lysine.

The transition enthalpy deltaH of the helix-random coil transition of the DNA-polylysine complex was measured as a function of the peptide:nucleotide ratio by the help of an adiabatic scanning differential calorimeter. Furthermore the transition enthalpy of a complex with a specific peptide:nucleotide ratio was determined as a function of the cation concentration of the solution. Finally the reaction enthalpy of the interaction of polylysine with native and denatured DNA was measured with the help of a LKB batch calorimeter. From the results of the calorimetric measurements one can conclude that the transition enthalpy of the DNA-polylysine complexes is linearly dependent on the nucleotide: peptide ratio. The extrapolated value for the 1:1 complex is 14.4 kcal per mole base pairs.

Animals↗

[A calorimetric study of the triple helix formation: interaction of poly(C) - guanosine - protonated poly(C)].

The triple helix formation of poly(C) - guanosine - poly(C+) was investigated by the help of an LKB scanning micro-calorimeter. The existence of the triple helix could also be shown by recording the melting curves. The ultraviolet absorption at different wave lengths namely 275 nm, 260 nm, and 245 nm was plotted as a function of the temperature. Furthermore formation of the triple helix was shown by plotting the ultraviolet absorption at 245 nm during the increasing addition of guanosine solution to a fixed amount of poly(C) in the solution. Finally the formation of the triple helix was demonstrated by plotting the ultraviolet absorption at 245 nm of a certain mixture of the components while the pH value of the solution was continuously lowered. All these methods show that the monomer interacts with the polymer double helix to form a triple helix. The calorimetric measurements show that the reaction enthalpy is concentration dependent. Above a threshold concentration a rapid increase of the reaction enthalpy is observed. This increase occurs in a very narrow concentration interval. Above this interval a final value of the reaction enthalpy is reached. The amount of the reaction enthalpy for the interaction of guanosine with poly(C) - poly(C+) double helix is 5.5 Kcal (mol base triplet)-1.

Calorimetry↗