PubMed Health⌕ Search

Biomedical subjects

H Hinz

Publications and source records attributed to H Hinz.

12 recordsLinked to original sources
PubMed2002

A review and assessment of tributyltin contamination in the North Sea, based on surveys of butyltin tissue burdens and imposex/intersex in four species of neogastropods.

It is evident from measures of butyltin tissue burdens and imposex or intersex in neogastropods that tributyltin (TBT) contamination of coastal waters and open parts of the North Sea is now low. It has been declining for at least the past decade. This is probably due to two measures. First, regulations prohibiting the use of TBT-based paints on small boats and fish farms have reduced inputs of TBT from these sources so that they are now negligible (except possibly where the regulations are flaunted). Second, there is evidence from sites, where commercial vessels are the sole source of TBT, that the adoption of TBT SPC paints has been effective in reducing environmental levels of these contaminants. However, poor dockyard practices, allowing TBT-contaminated wastes, including paint flakes, to accumulate in sediments have left a legacy of hot-spots of contamination in some ports. The impact is localised so that TBT contamination is low in coastal areas immediately adjacent to ports.

Animals↗
PubMed2001

Human micro-calpain: simple isolation from erythrocytes and characterization of autolysis fragments.

Heterodimeric p-calpain, consisting of the large (80 kDa) and the small (30 kDa) subunit, was isolated and purified from human erythrocytes by a highly reproducible four-step purification procedure. Obtained material is more than 95% pure and has a specific activity of 6-7 mU/mg. Presence of contaminating proteins could not be detected by HPLC and sequence analysis. During storage at -80 degrees C the enzyme remains fully activatable by Ca2+, although the small subunit is partially processed to a 22 kDa fragment. This novel autolysis product of the small subunit starts with the sequence 60RILG and is further processed to the known 18 kDa fragment. Active forms and typical transient and stable autolysis products of the large subunit were identified by protein sequencing. In casein-zymograms only the activatable forms 80 kDa+30 kDa, 80 kDa+22 kDa and 80 kDa+18 kDa displayed caseinolysis.

Autolysis↗
PubMed2000

A new alternative method to quantify residual structure in 'unfolded' proteins.

Pig (pCSD1) and human (hCSD1) calpastatin domain 1 proteins were studied to characterize common features of the denatured state of proteins. These proteins were chosen for the present investigation, because pCSD1 was suggested previously to be unstructured in water even at 25 degrees C (1) [T. Konno et al., Biochim. Biophys. Acta 1342 (1997) 73-82]. hCSD1 could be expected to exhibit similar features on the basis of preliminary spectroscopic studies. In the present study, the experimental grounds for the estimate of residual structure in the unfolded state were differential scanning calorimetry heat capacity and circular dichroism (CD) measurements over the temperature range 10-80 degrees C. At selected temperatures, we studied also the effect of guanidinium hydrochloride (GdnHCl) which is known to promote further unfolding of the polypeptide chain. All other measurements were performed at pH 6 in pure water. The present results support the conclusion that the comparison of the experimentally obtained heat capacity data with theoretical heat capacity values calculated on the basis of a newly established increment system gives insight into the degree of hydration of the unfolded polypeptide chain. The percentage by which the experimental heat capacity of the unfolded polypeptide chain differs from the calculated heat capacity permits a quantitative estimate of the residual structure. This estimate is in good agreement with that based on CD absorption. The heat capacity approach has the advantage of comparing fully hydrated and partially hydrated residues in the same aqueous environment, whereas for example spectroscopic measurements, such as CD, are generally referred to the fully unfolded chain in concentrated urea or GdnHCl solutions. As the unfolded chains of pCSD1 and hCSD1 exhibit a smaller heat capacity than that calculated on the new peptide-based increment system [M. Häckel et al., J. Mol. Biol. 291 (1999) 197-213], we conclude that the residues in the unfolded polypeptide chain are less hydrated than the same residues in oligopeptides. This suboptimal hydration is the result of residual structure in the chain as observed in both CD and heat capacity measurements.

Amino Acid Sequence↗
PubMed1999

Not more than three tissue kallikreins identified from organs of the guinea pig.

The large and varied multigene families of tissue kallikreins of rat and mouse are considered to selectively release as many bioactive peptides. In order to determine whether a similar family of enzymes is expressed in the organs of the guinea pig purification studies were performed. Tissue kallikreins from the submandibular gland, coagulating gland/prostate complex and the pancreas were separated by affinity chromatography on benzamidine-Sepharose. Amino-terminal sequences, the patterns of hydrolysis rates of a number of peptide p-nitroanilides, inactivation rates by active site-directed irreversible inhibitors, specific kininogenase activities and types of kinin released were used to probe the identity of the isolated enzymes. Guinea pig tissue kallikreins 1 and 2 have been reported previously. In the present study we have identified a third type, designated tissue kallikrein 1a because of its sequence similarity to kallikrein 1, which differs from the latter in the catalytic properties. The inferred occurrence of not more than two or three independent tissue kallikrein genes in the guinea pig contrasts with the varied family of enzymes expressed by the large number of such genes present in rats and mice. Expression in the guinea pig (and also in humans) of only a small number of tissue kallikreins makes specific processing of a multitude of biologically active peptides by such enzymes unlikely.

Amino Acid Sequence↗
PubMed1994

No intermediate channelling in stepwise hydrolysis of fluorescein di-beta-D-galactoside by beta-galactosidase.

For the hydrolysis of the two glycosidic bonds of fluorescein di-beta-D-galactoside (FDG) by beta-galactosidase from Escherichia coli, small [Hofmann, J. & Sernetz, M. (1983) Anal. Biochem. 131, 180-186] to dramatic [Huang, Z. (1991) Biochemistry 30, 8535-8540] deviations from simple stepwise substrate-intermediate-product kinetics have been reported. Intermediate channelling, a preferred hydrolysis of the intermediate fluorescein mono-beta-D-galactoside (FMG) formed from FDG at the active site and thus in a favourable position for further reaction, has been postulated. As there were reasons to doubt the previous findings and conclusions, the hydrolysis experiments have been repeated at initial FDG concentrations of 7-200 microM, following the concentrations of FDG, FMG and fluorescein with a reliable method, quantitative HPLC, to completion of the reaction. The transient appearance of substantial amounts of the intermediate FMG also in experiments with 200 microM FDG already rules out the existence of the most efficient intermediate channelling deduced by Huang (1991) from measurements of the initially developing fluorescence, incorrectly ascribed to fluorescein. Redetermination of the Michaelis constants for FDG and FMG led to much higher values than those reported previously. Fitting the progress curves by means of nonlinear regression combined with numerical integration of the rate equations resulted in good fits of the normal stepwise substrate-intermediate-product mechanism, without any necessity of assuming a more complex course of the reaction. So one of the rare examples of the hydrolysis of two bonds at a single enzyme-substrate encounter has been invalidated.

Chromatography, High Pressure Liquid↗
PubMed1992

Kinetics of bond cleavages at kallidin release by tissue kallikrein: cleavage of two peptide bonds in a single enzyme-substrate complex?

The kinetics of the release of kallidin, L- and KL-chains from bovine L-kininogen by porcine tissue kallikrein were followed and individual kinetic constants for cleavage of the Met-360 and the Arg-370 bond determined. The results suggest that both these bonds in L-kininogen r are hydrolyzed "simultaneously" without appearance of a free singly-nicked intermediate. Kallidin release in the human analogous system is also compatible with such a mechanism.

Animals↗
PubMed1991

Camptothecin and its derivatives induce expression of the c-jun protooncogene in human myeloid leukemia cells.

We have recently demonstrated that certain camptothecin derivatives are effective agents in the treatment of human tumor xenografts in nude mice. While camptothecin and its derivatives are recognized as inhibitors of topoisomerase I, little is known about the effects of these agents on specific gene expression, particularly genes involved in growth control. The c-jun early response gene codes for a leucine zipper transcription factor. The present studies demonstrate that 20(S)-camptothecin, 9-amino-20(S)-camptothecin, and 9-nitro-20(S)-camptothecin inhibit the growth of human U-937 myeloid leukemia cells and induce expression of the c-jun gene. c-jun transcripts were increased at 3 h and reached a maximum at 6 h of drug exposure. We also demonstrate that the induction of c-jun gene expression by these agents occurs at the transcriptional level. H7, a nonselective inhibitor of protein kinase C, completely blocked c-jun expression in 20(S)-camptothecin-treated cells, while another protein kinase inhibitor, HA1004, had no detectable effect. Similar findings were obtained for other leucine zipper encoding genes, including jun-B. These results suggest that 20(S)-camptothecin, 9-amino-20(S)-camptothecin, and 9-nitro-20(S)-camptothecin activate a cellular response involving the induction of early response genes. Finally, we demonstrate that induction of c-jun expression occurs in association with internucleosomal DNA fragmentation, a characteristic of programmed cell death.

Camptothecin↗
PubMed1989

[Projective identification and the psychoanalytic dialogue].

A survey of several psychoanalytic conceptions of projective identification is followed by a discussion, with case illustrations, of its communicative and defensive function. In this context, special attention is paid to the use of language as an instrument for the destruction of the object--in this case, the analyst--who is simultaneously yearned for and experienced as a threat.

Adult↗
PubMed1986

Individual reaction steps in the release of kallidin from kininogen by tissue kallikrein.

At low pH values (around 6), porcine pancreatic beta-kallikrein B attacks at first the C-terminal ARg bond of the kinin moiety in bovine HMW kininogen. Arg-cleaved kininogen accumulates as an intermediate in the solution. Kallidin is released by cleavage of the aminoterminal Met-Lys bond in a second step. At pH values between 7.6 and 9, however, Arg-cleaved kininogen does not occur as a free intermediate. The participation as a (free, not only enzyme-bound) intermediate of Arg-cleaved kininogen in a short-lived especially reactive conformation or of Met-cleaved kininogen is also unlikely. Probably, both the Met and the Arg bonds are hydrolyzed in one enzyme-substrate complex which does not dissociate between these two events. Kinetic constants for the release of kallidin from native single-chain HMW kininogen and from Arg-cleaved kininogen (even if this Arg residue is removed) remarkably have the same values. Evidently, the rate of the reaction is determined by steps leading to the hydrolysis of the Met bond. As the state of the C-terminal Arg residue has no influence, the efficient cleavage of the Met bond by tissue kallikrein is probably not due to some strain in the kininogen molecule in the region of this bond. As modification of Arg residues of kininogen prevents cleavage also of the Met bond, some Arg residue(s) appear(s) to play a crucial role in this process. kcat/Km (1.4 x 10(6) M-1 sex-1 at pH 9, 25 degrees C) is very high for a proteolytic reaction, mainly because of the low value of Km (0.6 microM).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗