| 41. |
- Abrahamson, Magnus, et al.
(författare)
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Identification of the probable inhibitory reactive sites of the cysteine proteinase inhibitors human cystatin C and chicken cystatin
- 1987
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Ingår i: Journal of Biological Chemistry. - 1083-351X. ; 262:20, s. 9688-9694
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Tidskriftsartikel (refereegranskat)abstract
- When an excess of human cystatin C or chicken cystatin was mixed with papain, an enzyme-inhibitor complex was formed immediately. The residual free cystatin was then progressively converted to a form with different electrophoretic mobility and chromatographic properties. The modified cystatins were isolated and sequenced, showing that there had been cleavage of a single peptide bond in each molecule: Gly11-Gly12 in cystatin C, and Gly9-Ala10 in chicken cystatin. The residues Gly11 (cystatin C) and Gly9 (chicken cystatin) are among only three residues conserved in all known sequences of inhibitory cystatins. The modified cystatins were at least 1000-fold weaker inhibitors of papain than the native cystatins. An 18-residue synthetic peptide corresponding to residues 4-21 of cystatin C did not inhibit papain but was cleaved at the same Gly-Gly bond as cystatin C. When iodoacetate or L-3-carboxy- trans-2,3-epoxypropionyl-leucylamido-(4-guanidin o)butane was added to the mixtures of either cystatin with papain, modification of the excess cystatin was blocked. Papain-cystatin complexes were stable to prolonged incubation, even in the presence of excess papain. We conclude that the peptidyl bond of the conserved glycine residue in human cystatin C and chicken cystatin probably is part of a substrate- like inhibitory reactive site of these cysteine proteinase inhibitors of the cystatin superfamily and that this may be true also for other inhibitors of this superfamily. We also propose that human cystatin C and chicken cystatin, and probably other cystatins as well, inhibit cysteine proteinases by the simultaneous interactions with such proteinases of the inhibitory reactive sites and other, so far not identified, areas of the cystatins. The cleavage of the inhibitory reactive site glycyl bond in mixtures of papain with excess quantities of cystatins is apparently due to the activity of a small percentage of atypical cysteine proteinase molecules in the papain preparation that form only very loose complexes with cystatins under the conditions employed and degrade the free cystatin molecules.
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| 42. |
- Abrahamson, Magnus, et al.
(författare)
-
Increased body temperature accelerates aggregation of the (Leu-68–>Gln) mutant cystatin C, the amyloid-forming protein in hereditary cystatin C amyloid angiopathy
- 1994
-
Ingår i: Proceedings of the National Academy of Sciences. - 1091-6490. ; 91:4, s. 1416-1420
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Tidskriftsartikel (refereegranskat)abstract
- Hereditary cystatin C amyloid angiopathy is a dominantly inherited disorder, characterized by dementia, paralysis, and death from cerebral hemorrhage in early adult life. A variant of the cysteine proteinase inhibitor, cystatin C, is deposited as amyloid in the tissues of the patients and their spinal-fluid level of cystatin C is abnormally low. The disease-associated Leu-68-->Gln mutant (L68Q) cystatin C has been produced in an Escherichia coli expression system and isolated by use of denaturing buffers, immunosorption, and gel filtration. Parallel physicochemical and functional investigations of L68Q-cystatin C and wild-type cystatin C revealed that both proteins effectively inhibit the cysteine proteinase cathepsin B (equilibrium constants for dissociation, 0.4 and 0.5 nM, respectively) but differ considerably in their tendency to dimerize and form aggregates. While wild-type cystatin C is monomeric and functionally active even after prolonged storage at elevated temperatures, L68Q-cystatin C starts to dimerize and lose biological activity immediately after it is transferred to a nondenaturing buffer. The dimerization of L68Q-cystatin C is highly temperature-dependent, with a rise in incubation temperature from 37 to 40 degrees C resulting in a 150% increase in dimerization rate. The aggregation at physiological concentrations is likewise increased at 40 compared to 37 degrees C, by approximately 60%. These properties of L68Q-cystatin C have bearing upon our understanding of the pathophysiological process of hereditary cystatin C amyloid angiopathy. They might also be of clinical relevance, since medical intervention to abort febrile periods of carriers of the disease trait may reduce the in vivo formation of L68Q-cystatin C aggregates.
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| 43. |
- Abrahamson, Magnus, et al.
(författare)
-
Isolation of six cysteine proteinase inhibitors from human urine. Their physicochemical and enzyme kinetic properties and concentrations in biological fluids
- 1986
-
Ingår i: Journal of Biological Chemistry. - 1083-351X. ; 261:24, s. 11282-11289
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Tidskriftsartikel (refereegranskat)abstract
- Six cysteine proteinase inhibitors were isolated from human urine by affinity chromatography on insolubilized carboxymethylpapain followed by ion-exchange chromatography and immunosorption. Physicochemical and immunochemical measurements identified one as cystatin A, one as cystatin B, one as cystatin C, one as cystatin S, and one as low molecular weight kininogen. The sixth inhibitor displayed immunochemical cross-reactivity with salivary cystatin S but had a different pI (6.85 versus 4.68) and a different (blocked) N-terminal amino acid. This inhibitor was tentatively designated cystatin SU. The isolated inhibitors accounted for nearly all of the cysteine proteinase inhibitory activity of the urinary pool used as starting material. The enzyme inhibitory properties of the inhibitors were investigated by measuring inhibition and rate constants for their interactions with papain and human cathepsin B. Antisera raised against the inhibitors were used in immunochemical determinations of their concentrations in several biological fluids. The combined enzyme kinetic and concentration data showed that several of the inhibitors have the capacity to play physiologically important roles as cysteine proteinase inhibitors in many biological fluids. Cystatin C had the highest molar concentration of the inhibitors in seminal plasma, cerebrospinal fluid, and milk; cystatin S in saliva and tears; and kininogen in blood plasma, synovial fluid, and amniotic fluid.
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| 44. |
- Abrahamson, Magnus, et al.
(författare)
-
Molecular cloning and sequence analysis of cDNA coding for the precursor of the human cysteine proteinase inhibitor cystatin C
- 1987
-
Ingår i: FEBS Letters. - : Wiley. - 1873-3468 .- 0014-5793. ; 216:2, s. 229-233
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Tidskriftsartikel (refereegranskat)abstract
- Recombinant cystatin C producing clones were isolated from a human placenta λgt11 cDNA library. The cDNA insert of one of the clones, containing 777 base pairs, encodes the complete mature cystatin C (120 amino acids) and a hydrophobic leader sequence of 26 amino acids, indicating an extracellular function of the inhibitor. The deduced protein sequence confirms the protein sequence of cystatin C isolated from human urine, but differs in one position from the sequence of the cystatin C fragment deposited as amyloid in hereditary cerebral hemorrhage with amyloidosis.
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| 45. |
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| 46. |
- Abrahamson, Magnus, et al.
(författare)
-
Structure and expression of the human cystatin C gene
- 1990
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Ingår i: Biochemical Journal. - 1470-8728. ; 268:2, s. 287-294
-
Tidskriftsartikel (refereegranskat)abstract
- The structural organization of the gene for the human cysteine-proteinase inhibitor cystatin C was studied. Restriction-endonuclease digests of human genomic DNA hybridized with human cystatin C cDNA and genomic probes produced patterns consistent with a single cystatin C gene and, also, the presence of six closely related sequences in the human genome. A 30 kb restriction map covering the genomic region of the cystatin C gene was constructed. The positions of three polymorphic restriction sites, found at examination of digests of genomic DNA from 79 subjects, were localized in the flanking regions of the gene. The gene was cloned and the nucleotide sequence of a 7.3 kb genomic segment was determined, containing the three exons of the cystatin C structural gene as well as 1.0 kb of 5'-flanking and 2.0 kb of 3'-flanking sequences. Northern-blot experiments revealed that the cystatin C gene is expressed in every human tissue examined, including kidney, liver, pancreas, intestine, stomach, antrum, lung and placenta. The highest cystatin C expression was seen in seminal vesicles. The apparently non-tissue-specific expression of this cysteine-proteinase inhibitor gene is discussed with respect to the structure of its 5'-flanking region, which shares several features with those of housekeeping genes.
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| 47. |
- Abrahamson, Magnus, et al.
(författare)
-
The human cystatin C gene (CST3), mutated in hereditary cystatin C amyloid angiopathy, is located on chromosome 20
- 1989
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Ingår i: Human Genetics. - 1432-1203. ; 82:3, s. 223-226
-
Tidskriftsartikel (refereegranskat)abstract
- Hereditary cystatin C amyloid angiopathy has recently been shown to be caused by a point mutation in the cystatin C gene. To determine the chromosomal localization of the gene, 20 human-rodent somatic cell hybrids and a fulllength cystatin C cDNA probe were used. Southern blot analysis of BamHI digested cell hybrid DNA revealed that the probe recognizes a 10.6 kb human specific fragment and that this fragment cosegregates with human chromosome 20. Therefore, the human cystatin C gene (CST3) was assigned to chromosome 20.
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| 48. |
- Abrahamsson, Bengt
(författare)
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Det gick som det gick. Om inre logik, särskilt i organisationer
- 1994
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Ingår i: Sociologisk forskning. - : Sveriges Sociologförbund. - 0038-0342 .- 2002-066X. ; 31:3, s. 3-22
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Tidskriftsartikel (refereegranskat)abstract
- Whatever happened, happened. Some notes on inner logic, especially in organizationsThe notion that social events partly arise as a consequence of inner logic, i.e. that patterns and structures emerge outside of, or even in opposition against, plans and goals is a common element in social science. Inner logic is a summary term for social processes developing autonomously, i.e. without any individual or group intending them. Organizations quite often contain inner-logic processes. If, as this author maintains, fruitful organization theory has to build on rationalistic assumptions, how then do we handle instances of inner logic? A first step may be to break up the traditional link between structuralism and functionalism, maintaining the former and rejecting the latter. Organizations are intentionally dynamic, i.e. depend on order and predictability. To the extent that inner-logic processes appear in organizations, they should be analysed as confrontations between opposing rationalities rather than as spontaneous reactions of a ”system” . Also, frequently recurring organizational forms such as hierarchy are more fruitfully regarded as e.g. transaction-cost outcomes rather than as functional responses to system needs. Rationalism and structuralism are compatible, rationalism and functionalism are not.
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| 49. |
- Abrahamsson, Hans, 1966, et al.
(författare)
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A Turbulent Plane 2-Dimensional Wall-Jet in a Quiescent Surrounding
- 1994
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Ingår i: European Journal of Mechanics/B-Fluids. - 0997-7546 .- 1873-7390. ; 13:5, s. 533-556
-
Tidskriftsartikel (refereegranskat)abstract
- An experimental investigation of the turbulence field in a plane two-dimensional wall-jet has been conducted. All measurements were carried out employing hot-wire techniques in a large scale experimental facility. Mean velocities, Reynolds stresses and wall shear stress were determined with slot Reynolds numbers of 1.0*104, 1.5*104 and 2.0*104. The wall-jet was found to be self-preserving, and an asymptotic scale matching was used to show the existence of a short inertial sub layer as compared with a wall boundary layer. Studies of the integral scale development and the turbulent diffusion of shear stress showed a strong interaction between the inner and outer regions of the wall-jet. Using momentum scaling, it was found that the streamwise development of the half-width and maximum mean velocity was independent of the slot Reynolds number.
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| 50. |
- Abrahamsson, Hans, 1966, et al.
(författare)
-
Turbulence in a Two-Dimensional Wall-Jet - Experiments and Modeling
- 1992
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Ingår i: 13th Turbulence Conference in Missouri-Rolla..
-
Konferensbidrag (refereegranskat)abstract
- An experimental and numerical investigation of the turbulence field in a two-dimensional wall-jet has been carried out in a well-defined geometry. The measurements were performed using a hot-wire technique and the calculations were carried out with a standard k-ε model as well as an algebraic stress model (ASM), using elliptic solvers. For the mean velocity field, good agreements were found between the measurements and the calculations. The measurements of the turbulence intensities reveal two maxima in the stream-wise and span-wise components, while only one maximum was found in the normal component. Also in the shear stress two maxima were found. A comparison between calculations and measurements indicates that improved turbulence models are needed.
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