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B Hollenbach

Publications and source records attributed to B Hollenbach.

6 recordsLinked to original sources

Aggregation of truncated GST-HD exon 1 fusion proteins containing normal range and expanded glutamine repeats.

We have shown previously by electron microscopy that the purified glutathione S-transferase (GST)-Huntington's disease (HD) exon 1 fusion protein with 51 glutamine residues (GST-HD51) is an oligomer, and that site-specific proteolytic cleavage of this fusion protein results in the formation of insoluble more highly ordered protein aggregates with a fibrillar or ribbon-like morphology (E. Scherzinger et al. (1997) Cell 90, 549-558). Here we report that a truncated GST HD exon 1 fusion protein with 51 glutamine residues, which lacks the proline-rich region C-terminal to the polyglutamine (polyQ) tract (GST-HD51 delta P) self-aggregates into high-molecular-mass protein aggregates without prior proteolytic cleavage. Electron micrographs of these protein aggregates revealed thread-like fibrils with a uniform diameter of ca. 25 nm. In contrast, proteolytic cleavage of GST-HD51 delta P resulted in the formation of numerous clusters of high-molecular-mass fibrils with a different, ribbon-like morphology. These structures were reminiscent of prion rods and beta-amyloid fibrils in Alzheimer's disease. In agreement with our previous results with full-length GST-HD exon 1, the truncated fusion proteins GST-HD20 delta P and GST-HD30 delta P did not show any tendency to form more highly ordered structures, either with or without protease treatment.

Cloning, Molecular↗

Huntingtin aggregation monitored by dynamic light scattering.

An initial stage of fibrillogenesis in solutions of glutathione S-transferase-huntingtin (GST-HD) fusion proteins has been studied by using dynamic light scattering. Two GST-HD systems with poly-L-glutamine (polyGln) extensions of different lengths (20 and 51 residues) have been examined. For both systems, kinetics of z-average translation diffusion coefficients (Dapp) and their angular dependence have been obtained. Our data reveal that aggregation does occur in both GST-HD51 and GST-HD20 solutions, but that it is much more pronounced in the former. Thus, our approach provides a powerful tool for the quantitative assay of GST-HD fibrillogenesis in vitro.

Glutathione Transferase↗

Huntingtin-encoded polyglutamine expansions form amyloid-like protein aggregates in vitro and in vivo.

The mechanism by which an elongated polyglutamine sequence causes neurodegeneration in Huntington's disease (HD) is unknown. In this study, we show that the proteolytic cleavage of a GST-huntingtin fusion protein leads to the formation of insoluble high molecular weight protein aggregates only when the polyglutamine expansion is in the pathogenic range. Electron micrographs of these aggregates revealed a fibrillar or ribbon-like morphology, reminiscent of scrapie prions and beta-amyloid fibrils in Alzheimer's disease. Subcellular fractionation and ultrastructural techniques showed the in vivo presence of these structures in the brains of mice transgenic for the HD mutation. Our in vitro model will aid in an eventual understanding of the molecular pathology of HD and the development of preventative strategies.

Amino Acid Sequence↗

Cadmium leads to stimulated expression of the lipid transfer protein genes in barley: implications for the involvement of lipid transfer proteins in wax assembly.

In order to investigate the nature of genes expressed in leaf epidermal cells of higher plants, we have identified the nucleotide sequence of a cDNA designated ltp 7a2b encoding a novel nonspecific lipid transfer protein of barley (Hordeum vulgare L. cv. Gerbel). The cDNA of 755 basepairs contains an open reading frame of 366 nucleotides coding for a 12.3-kDa polypeptide. The first 29 amino acids constitute the putative signal peptide, characteristic for targeting to the secretory pathway. Analysis of mRNA levels by Northern blotting indicated that ltp 7a2b is preferentially expressed in the leaf epidermis. Levels of mRNA decreased during ageing of leaf tissue. Expression of ltp 7a2b was stimulated by a factor of 2 - 3 when the seedlings were grown in the presence of cadmium (10 - 1600 microM). Concomitantly, the primary leaves of Cd-exposed seedlings contained elevated levels of abscisic acid and thicker was layer of the cuticle. At 100 microM Cd in the hydroponic medium, the was cover was increased by 50%. The increase in abscisic acid content, ltp 7a2b mRNA and was coverage was either not seen, or seen much less, in Ni- and Zn-stressed seedlings. The data add circumstantial evidence to the recently proposed hypothesis that nonspecific lipid transfer proteins function in transfer of cutin and/or wax monomers from the site of synthesis in the cell to the cuticle.

Amino Acid Sequence↗

[Pregnancy, labour, and puerperium in heroin addicted women, with reference to experience and the present state of knowledge (author's transl)].

The mounting problems of drug addiction now also confront the obstetrician and pediatrician. 1/3 of the 60,000 addicts to opiates in the Federal Republic of Germany are women. Of these 80% are in the reproductive age group. Despite endocrine problems pregnancies do occur and are high risk pregnancies because of hepatitis, venereal disease, malnutrition, phlebitis, abscesses, premature deliveries, premature rupture of the membranes, malpresentations, dysmaturity, pre-eclampsia, and numerous other social and psychological problems. Because of the varying content of heroin on the black market the fetus runs the risk of acute overdose or withdrawal. Withdrawal results in extremely marked fetal movements, with increased oxygen consumption and a danger of intra-uterine asphyxia sometimes resulting in intra-uterine fetal death. Most newborns develop a withdrawal syndrome with irritability, high pitched cries, shivering, tachycardia, perspirations, fever and generalized seizures. The experience of the Department for Women for the Free University in Berlin-Charlottenburg are presented and compared to a review of the literature. The medical, social, and psychological aspects of the problem are discussed.

Female↗