Search PubMed⌕ Search

Biomedical subjects

J Schaller

Publications and source records attributed to J Schaller.

At least 55 records · Page 3Linked to original sources
PubMed1996

Identification of disulfide bonds in the ninth component (C9) of human complement.

C9 is the most abundant protein of the membrane attack complex of complement. By means of limited proteolysis, different chromatographic techniques, a thiol-specific fluorescence assay, amino acid analysis, and Edman degradation 9 out of 12 disulfide bridges are definitely assigned (Cys22-Cys57, Cys33-Cys36, Cys67-Cys73, Cys121-Cys160, Cys233- Cys234, Cys359-Cys384, Cys489-Cys505, Cys492-Cys507, Cys509-Cys518). Weaker evidence permits to reduce the number of possible configurations for the remaining 3 cystines (Cys80-Cys91, Cys86-Cys104, Cys98-Cys113, or Cys80-Cys91, Cys86-Cys113, Cys98-Cys104). These findings are discussed in comparison with the strongly related components C6, C7, C8alpha, and C8beta.

Amino Acid Sequence↗
PubMed1996

[HPV-5 typing with nested PCR and sequencing in epidermodysplasia verruciformis].

We report a case of epidermodysplasia verruciformis (EV) with typical cutaneous manifestations in light-exposed areas. The diagnostic criteria were the characteristic macular lesions, the virus transformed keratinocytes on histological examination and the detection of human papillomavirus (HPV) DNA by a nested-PCR approach for EV-associated types. In this case we found HPV-type 5, the most common type of HPV in EV, directly by sequencing and sequence analysis.

Adult↗
PubMed1996

[Cutaneous malacoplakia in a patient with psoriasis vulgaris].

A 58 year old patient suffering from psoriasis vulgaris had typical erythematous, scaly plaques for 20 years. In addition, an erosive, therapy-resistant plaque was found in the perianal region. A biopsy was taken to exclude malignancy; it showed intracytoplasmic Michaelis-Gutmann-bodies as a marker for malakoplakia, which rarely occurs in the skin. In further electron microscopic investigation, intracytoplasmatic bacteria could be demonstrated. An underlying malignancy or chronic renal insufficiency with immunosuppression were not found in this patient; both are often associated with malakoplakia.

Anal Canal↗
PubMed1996

Low expression of adhesion molecules and matrix proteins in patients showing poor penetration in zona-free hamster oocytes.

The expression of adhesion molecules and matrix proteins by human spermatozoa as well as their binding to and penetration of zona-free hamster eggs were investigated in 17 patients by means of flow cytometry. Both binding and penetration of hamster oocytes and expression of alpha- and beta-chains of beta 1, beta 3 and beta 4 integrins were determined before and after induction of the acrosome reaction. The expression of the integrin ligands, fibronectin and laminin were also analysed. Significant differences in the expression of very late antigen (VLA) integrins, VLA alpha 4-chain (CD49d), the classical fibronectin receptor VLA alpha 5-chain (CD49e), leukocyte function-associated molecule-3 (LFA-3; CD58) and fibronectin were observed between patients showing good (> 10%) or poor (< 10%) penetration in the sperm penetration assay (P = 0.0068). It is concluded that these adhesion molecules are intimately involved in the sperm-oolemma interaction. Since no differences were observed in either spermiogram parameters or the percentages of acrosome-reacted spermatozoa in the two groups, sperm-oolemma binding and penetration should be regarded as discrete parameters of sperm function.

Animals↗
PubMed1996

Detection of mRNA transcripts of beta 1 integrins in ejaculated human spermatozoa by nested reverse transcription-polymerase chain reaction.

Matrix proteins (e.g. fibronectin, laminin and vitronectin) and their receptors, especially beta 1 integrins, may play a role in sperm-oocyte interactions. These proteins are well described on spermatogenic cells but especially in ejaculated spermatozoa, the presence of mRNA as a necessity for their synthesis is not clear. Therefore the mRNA synthesis of the beta 1, alpha 3, alpha 4, alpha 5 and alpha 6 chains (fibronectin and laminin receptors) of the beta 1 integrins was investigated by nested reverse transcription-polymerase chain reaction (RT-PCR) in spermatozoa of human semen samples (n = 18) which showed normal classical semen parameters after swim-up preparation. After the second amplification, we obtained specific signals for mRNA transcripts of the beta 1, alpha 3, alpha 4 and alpha 6 chains in 17 of 18 semen samples investigated. All control PCRs were negative for somatic cell transcripts and contamination by somatic cells was excluded. These results demonstrate for the first time that human ejaculated spermatozoa contain the mRNA transcripts of beta 1 integrins and support the role of these proteins in adhesive mechanisms in the fertilization process.

Actins↗
PubMed1995

Partial amino acid sequence and functional aspects of histone H1 proteins in Trypanosoma brucei brucei.

Trypanosoma brucei brucei, a protozoan parasite of wild and domestic animals in Africa, is related to the pathogenic agent of human sleeping sickness. Four H1 histone proteins were isolated from nuclei of procyclic culture forms and cleaved with proteases. Amino acid sequence analysis of purified fragments indicated the presence of variants which displayed sequence identities as compared to the C-terminal domain of human H1. Substitutions of amino acids and posttranslational modifications of the histones in T b brucei H1 may influence protein conformation and histone-histone as well as histone-DNA interactions in the chromatin of the parasite. Digestion of soluble chromatin with immobilized trypsin at low and high ionic strengths indicated an internal localization of H1 in the condensed chromatin. The influence of histone H1 of T b brucei on the compaction pattern of the chromatin was investigated by dissociation and reconstitution experiments. Electron microscopy revealed that trypanosome H1 was able to induce condensation of the chromatin of the parasite and of rat liver into dense tangles. After dephosphorylation of H1, 30 nm fibers were induced in rat liver chromatin, while the resulting fibers were distinctly thinner in T b brucei. It can be concluded that the absence of 30 nm fibers in T b brucei chromatin cannot be explained by the divergent variants and posttranslational phosphorylations of H1 only but rather by the influence of both, the divergent core histones, previously described, and H1 properties.

Amino Acid Sequence↗
PubMed1995

A method for monoclonal antibody isotype switching: anti-CD60 VH expression in a heavy chain-deficient hybridoma variant.

Alteration of monoclonal antibody isotype is desirable for a variety of purposes, including obtaining an improved reagent for investigative or therapeutic use. A variety of approaches for isotype switching, particularly from IgM to various IgG subclasses, have been described. Antibodies that recognize carbohydrate determinants on glycoproteins, glycolipids, or polysaccharides are generally of the IgM isotype. This includes all available antibodies to the human CD60 antigen, a determinant with cell coactivating properties described on a subset of T lymphocytes and on other cell types. In this report a new method for monoclonal antibody isotype switching is presented. A plasmid containing the VH regions of anti-CD60 linked to C gamma 1 was transfected into a spontaneously arising variant of the CD60 hybridoma that produced kappa light chain but no heavy chain. This transfected hybridoma line maintains stable production of useful quantities of IgG1 monoclonal anti-CD60 in vitro and in vivo.

Animals↗
PubMed1994

Coagulation factor XII Locarno: the functional defect is caused by the amino acid substitution Arg 353-->Pro leading to loss of a kallikrein cleavage site.

The dysfunctional coagulation factor XII (FXII) Locarno was purified from 2 L of the proposita's plasma. Studies to identify the molecular defect responsible for the lack of amidolytic and proteolytic activity of this FXII variant were performed. Amino acid sequence analysis of peptides obtained from FXII Locarno on activation with either trypsin or plasma kallikrein and dextran sulfate showed an amino acid substitution of Arg 353 by Pro. Thereby, the kallikrein cleavage site at Arg 353-Val 354 is lost. Although trypsin-activated FXII Locarno was fully cleaved at Arg 334-Asn 335 and at Arg 343-Leu 344, neither amidolytic nor proteolytic activity was generated. We conclude that proteolytic cleavage at Arg 343 in the absence of cleavage at Arg 353 is not sufficient to expose the enzymatic active site in FXII Locarno.

Amino Acid Sequence↗
PubMed1994

Enhanced cytotoxicity of amyloid beta-peptide by a complement dependent mechanism.

Amyloid beta-peptide (A beta) has been shown to activate the classical complement pathway in vitro. Here, we demonstrate that this interaction is fully capable of killing cells and damaging cellular processes in mixed hippocampal cultures from embryonic day 18 rat fetuses. Lactic acid dehydrogenase (LDH) release and morphologic changes were used to evaluate toxicity.

Amyloid beta-Protein Precursor↗
PubMed1994

The human 4F2 antigen: evidence for cryptic and noncryptic epitopes and for a role of 4F2 in human T lymphocyte activation.

T lymphocyte activation can be triggered through multiple distinct, but functionally related, pathways. Murine monoclonal antibodies (mAbs) have been used to characterize the surface components of several of these pathways, as well as structures whose function is not yet known. One such cell surface structure is the heterodimeric 4F2 antigen, which is expressed on activated and proliferating cells. Two new mAbs that recognize the heavy chain of the 4F2 antigen have been produced in our laboratory. One antibody, UM7F8, is comitogenic with soluble anti-CD2 and immobilized (but not soluble) anti-CD3 mAbs. The second antibody, termed UM2G12, appears to recognize a cryptic epitope on the 4F2 heavy chain and is not comitogenic for T cells. In view of the functional effects seen with UM7F8, and the highly regulated expression of the 4F2 antigen, it seems likely that 4F2 has a specific role in T cell development and activation.

Antibodies, Monoclonal↗
PubMed1994

Expression, purification and characterization of the recombinant kringle 2 and kringle 3 domains of human plasminogen and analysis of their binding affinity for omega-aminocarboxylic acids.

The kringle 2 (E161T/C162S/EEE[K2HPg/C169S]TT) and the kringle 3 (TYQ[K3HPg]DS) domains of human plasminogen (HPg) were expressed in Escherichia coli in an expression vector with the phage T5 promotor/operator element N250PSN250P29 and the cDNA sequence for a hexahistidine tail to facilitate the isolation of the recombinant protein. A coagulation factor Xa (FXa)-sensitive cleavage site was introduced to remove the N-terminal histidine tag. In r-K2, mutations E161T and C162S were introduced to enhance the FXa cleavage yield and C169S to replace the cysteine residue, participating in the inter-kringle disulfide bridge between kringles 2 and 3. Recombinant proteins were isolated by affinity chromatography on Ni(2+)-nitrilotriacetic acid/agarose and refolded under denaturing and reducing conditions followed by a non-denaturing and oxidising environment. The free thiol group in position 297 in r-K3 was selectively alkylated with iodoacetamide. The hexahistidine tail was successfully removed with FXa. The N-terminal sequence, the amino acid composition and the molecular mass analyses are in agreement with the expected data. The correct arrangement of the disulfide bonds was verified by sequence analysis of the corresponding thermolytic and subtilisin fragments. r-K2 exhibits weak binding to lysine-Bio-Gel. The weak binding affinity of r-K2 for omega-aminocarboxylic acids is confirmed by intrinsic fluorescence titration with 6-aminohexanoic acid (NH2C5COOH) indicating a Kd of approximately 401 microM. In contrast, r-K3 seems to be devoid of a binding affinity for omega-aminocarboxylic acids. Considering earlier determined Kd values of kringle 1, kringle 4 and kringle 5, the binding affinity of HPg kringle domains for NH2C5COOH is proposed to decrease in the following order, kringle 1 > kringle 4 > kringle 5 > kringle 2 > kringle 3.

Amino Acid Sequence↗
PubMed1994

[Value of the patient's own test substances in epicutaneous testing].

Patch testing with commercially available test kits of known allergens often leads to detection of the responsible allergen. In many cases, however, the responsible allergen cannot be found unless the patient's "own" products or chemicals are tested. An analysis of 2460 patch tests with patch test kits proposed by the DKG (German Contact Allergy Group) and a positive history of a patient's own substance showed positive results in 208 patients with 289 products. In 44% (129 out of 289) the responsible allergen could only be found by testing the patient's own substances, in 56% (160 out of 289) the same allergen as was responsible for the positive result with patients' endogenous substances was positive in one of the commercially available test kits. The substances that tested positive were medical products/adhesives (45%), cosmetics (39.4%), rubber materials (4.1%), leather materials (0.7%) and others (10%). Subsequent testing of the different components of the patients' own substances that tested positive was carried out in 45 cases and resulted in the detection of new allergens, such as tris-(2-hydroxyethyl)isocyanurate triacrylate. In 27 patients the endogenous substances had tested positive in the course of testing for occupational dermatitis, and in 10 of these positive testing of a patient's own product resulted in notification of an occupational dermatitis. These results show, that patch testing with the patients own products if very important for the finding of the responsible or even new allergens.

Adult↗
PubMed1994

[Detection of an unusual varicella zoster virus infection in an immunosuppressed patient with polymerase chain reaction].

We report a case of a 76-year-old patient who developed an atypical form of varicella zoster virus (VZV) reinfection. In addition to histological and serological confirmation of diagnosis, direct demonstration of VZV-DNA was possible by means of the nested polymerase chain reaction. Although VZV infection is usually diagnosed by clinical examination, the nested PCR is a sensitive and specific diagnostic procedure that can be useful especially in atypical cases.

Acyclovir↗
PubMed1994

Kringle solution structures via NMR: two-dimensional 1H-NMR analysis of horse plasminogen kringle 4.

The kringle 4 domain of equine plasminogen (ePgn/K4), a close variant of the human homolog (hPgn/K4), contains residues, such as Trp32, which also appear in human apolipoprotein(a) kringle 4-type modules. The ePgn/K4 was investigated as a complex with epsilon-aminocaproic acid, an antifibrinolytic drug, by two-dimensional 1H-NMR spectroscopy at 500 MHz. Secondary structure elements were recognized from sequential medium and long-range dipolar (proton Overhauser) interactions, as well as from the identification of resonances originating from backbone amide protons with slow 1H-2H exchange in 2H2O. Antiparallel beta-sheets, consisting of strands 52-53, 61-65 and 71-75, were identified. Additionally, the segments 14-16 and 20-22 were found to assume characteristic interstrand antiparallel (beta-sheet-like) H-bond pairing. Four type I turns could be identified in strands 6-9, 16-19, 24-27 and 67-70. Ten structures were generated using distance geometry methods, followed by dynamic simulated annealing calculations. The root mean squares deviation of the distances was 2.79 A for all atoms and 1.81 A for backbone atoms only. Hydrogen bridges, involving side chain hydroxyl groups, were identified for Thr16 and Thr65. As observed for the hPgn/K4, the three-dimensional structure of the ePgn/K4 is mainly defined by two antiparallel beta-sheets, 14-16/20-22 and 62-66/71-75, which are oriented perpendicular to each other. Adjacent to these is a hydrophobic pocket, formed by Trp62, Tyr64, Trp72 and Phe74, whose side chains contribute a lipophilic component to the exposed lysine binding site surface. In contrast to the Trp25, Trp62 and Trp72 residues, conserved in the human and equine homologs, the spectrum of the Trp32 side chain reveals an unrestrained, solvent-exposed indole ring.

Amino Acid Sequence↗
PubMed1994

Purification of toxic peptides and the amino acid sequence of CSTX-1 from the multicomponent venom of Cupiennius salei (Araneae:Ctenidae).

The venom of the wandering spider Cupiennius salei was analysed biochemically by gel filtration, cation exchange chromatography, RP-HPLC, IEF, SDS-PAGE and TLC-electrophoresis. The native venom contains high levels of Na+, K+, Ca2+, histamine and taurine. It shows considerable activity of hyaluronidase, but not proteolytic activity. Thirteen peptides (CSTX-1 to CSTX-13) with an apparent mol. wt between 2.6 and 12.5 kDa causing differently strong toxic, effects were purified. Toxicity data of the crude venom (insects and mouse) are given and compared with the toxicity of CSTX-1, which causes most of the crude venom's toxicity. CSTX-1 has a mol. wt of 8352.6 and its amino acid sequence of 74 amino acids is given.

Amino Acid Sequence↗
PubMed1994

Identity of human extra parotid glycoprotein (EP-GP) with secretory actin binding protein (SABP) and its biological properties.

In this paper the identity of the salivary protein EP-GP (extra-parotid glycoprotein) is reported, also apparent in other human secretions. Immunochemical and biochemical analysis demonstrated that EP-GP is similar to the secretory actin-binding protein (SABP), also known as gross cystic disease fluid protein-15 (GCDFP-15) and prolactin-inducible protein (PIP). The molecular mass and charge microheterogeneity of EP-GP, also observed for SABP, was shown to be predominantly caused by the carbohydrate moiety. In addition, evidence was given that EP-GP is not related to the lipocalin Von Ebner's gland protein (human; VEGh). The biological significance of EP-GP and its homologues is not clear. EP-GP bound to actin and fibrinogen as described for SABP and GCDFP-15. However, the affinity for these proteins does not appear to have any direct physiological role in the mucosal secretions. On the other hand, EP-GP binds to several bacteria. By electron microscopy the ultrastructural localization is demonstrated of EP-GP to the cell wall of both Streptococcus salivarius HB and its cell appendage-lacking mutant Streptococcus salivarius HB-C12. Concerning this finding we hypothesize on the possible functional aspects of this enigmatic protein EP-GP.

Amino Acid Sequence↗