Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

---

Query: EC:3.4.21.4 (trypsin)
42,187 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Sections of formaldehyde-fixed paraffin-embedded cortical and hippocampal brain tissue from five cases with senile dementia of Alzheimer type (SDAT) and five cases with Pick's disease (PD) were immunostained with the monoclonal antibodies (mabs) 147, RT 97, BF 10 and 8D8 with and without pretreatment with alkaline phosphatase (AP) or trypsin (Tr). The mabs 147, RT 97 and BF 10 had previously been demonstrated to bind exclusively to phosphorylated epitopes of neurofilament proteins, while mab 8D8 is shown in this report to bind mainly, but not exclusively, to phosphorylated neurofilament epitopes. The mabs RT 97, BF 10 and 8D8, but not 147 stain most, if not all, Pick bodies (PB) and Alzheimer neurofibrillary tangles (NFT). When sections are pretreated with AP or Tr the immunostaining with mab BF 10 is very resistent in both PB and NFT. This resistance of PB and NFT is in contrast to the reduced staining of axons and of swollen cells in PD by the same enzymatic pretreatment. Immunostaining with mab RT 97 of PB and NFT is reduced moderately by AP and considerably by Tr. Only when stained with mab 8D8 is there a discrepancy between PB and NFT in their reaction to the pretreatment with AP: NFT staining with mab 8D8 is not affected, while that of PB is abolished. Thus, in spite of their different ultrastructure, PB and NFT are very similar immunocytochemically and in the accessibility of their phosphorylated epitopes to enzymatic treatment.
...
PMID:Alzheimer dementia and Pick's disease: neurofibrillary tangles and Pick bodies are associated with identical phosphorylated neurofilament epitopes. 244 59

Screening of antigenically reactive fragments of alpha S1-casein (alpha S1-CN), the major casein in bovine milk, was done by using HPLC and enzyme-linked immunosorbent assay (ELISA). BALB/c mice (6-week-old) were injected intraperitoneally with alpha S1-CN and complete Freund's adjuvant, and 14 days later, all the mice were boosted with alpha S1-CN and incomplete Freund's adjuvant. Twenty-one days after the 1st immunization, the mice were bled and antiserum was separated. Anti alpha S1-CN antibody fraction was obtained by precipitation from the antiserum with 50% saturated ammonium sulfate. alpha S1-CN was digested with trypsin and chymotrypsin, and 35 peptides were purified from the digests by reversed-phase HPLC with ODS (octadecylsilica) columns. Reactivity of peptides with the antibody were examined by ELISA. The solid phase in the wells of the polystyrene microtiter plate was coated with peptides, and the plate was successively incubated with anti alpha S1-CN antibody, conjugate of anti mouse immunoglobulin with alkaline phosphatase (ALP) and substrate of ALP. Two tryptic fragments (the residues 104-119 and 133-151) and three chymotryptic fragments (33-54, 105-121, and 174-199) were positive in an ELISA test. These five fragments would correspond to four antigenic sites. We could thus find antigenically reactive fragments of alpha S1-CN by the direct and simple detection of specific antigen-antibody interaction.
...
PMID:Rapid screening of antigenically reactive fragments of alpha s1-casein using HPLC and ELISA. 244 84

Use of prometaphase chromosome preparations has led to significant improvements in the localization of both NORs and ribosomal gene clusters in the short arms of human acrocentric chromosomes. An improvement of the NOR silver-staining method, followed by trypsin-Giemsa banding, was used to identify the precise location of the NOR on each human acrocentric chromosome. For comparison, the satellite, stalk, and centromeric region were also identified with the aid of both Q- and G-banding techniques. The amount of silver impregnation present in the stalk region of the D- and G-group chromosomes was unique for each of these acrocentric chromosomes and depended on the length of the stalk. In situ hybridization was used to locate the ribosomal gene clusters. A plasmid containing 5.6 kb of the 18S rDNA gene was first oligolabeled with bio-16-dUTP, then hybridized in situ to metaphase chromosomes and visualized by an alkaline phosphatase color-detection system. Our results indicated that, in most cases, the location of the 18S rDNA gene cluster in the stalk region was indistinguishable from the site of silver impregnation. However, exceptions were noted, suggesting a multiplicity of arrangements of the ribosomal gene clusters. A model is proposed to describe the spatial relationship of NORs (transcriptional activity) and the ribosomal gene clusters.
...
PMID:Visualization of NORs in relation to the precise chromosomal localization of ribosomal RNA genes. 247 78

The effect of matrix components extracted Bovine periodontal ligament (PDL) on cell proliferation and alkaline phosphatase activity of cultured human periodontal ligament fibroblast (HPLF) were examined in order to understand the cell-tissue interaction of periodontal ligament. Bovine PDL tissue was sequentially extracted with 0.05 M Tris HCl buffer, pH 7.4, containing 1M NaCl or 4M GdmCl. After seeding 24 hours, the cultured HPLF were exposed to the extracts for two through eight days. Nine days after seeding, HPLF indicated four times high activity on ALP and 1.6 times the amount of total protein than those of control (without extract), while DNA synthesis increased only 1.2 times. On the contrary, the NaCl extract depressed the ALP activity of HPLF. The GdmCl extract enhanced both the total protein and ALP activity in dose-dependently. The ALP increasing activity of GdmCl extract on HPLF is stable to heat (78 degrees C, 20 min) and collagenase treatment but partially inactivated by trypsin digestion. Since the GdmCl extract also induced colony formation of NRK-49F cell in soft agarose, it was suggested that the extract contain EGF and TGF-beta like factor. Molecular size of this factor was estimated as 20-50Kd using Sepharose CL-6B gel chromatography. Furthermore, this factor from Sepharose CL-6B were separated into two forms by ion exchange CM-Sepharose column chromatography. Purified preparation from reversed phase column chromatography contained 14Kd, 15Kd, 17Kd, 20Kd, 28Kd40Kd, and 46Kd components on SDS-PAGE. This factor may accumulate in extracellular matrix, and may play a role of cell-tissue interaction and homeostasis in periodontal ligament.
...
PMID:[Biochemical study on the TGF-beta like growth factor derived from bovine periodontal ligament]. 248 40

Saito et al recently reported that the alkaline phosphatase (ALPase) of human periodontal ligament fibroblasts (HPLF) showed remarkably high activity which was similar, but not identical phenotype, to that present in osteoblasts, and suggested that HPLF could be termed as "osteoblastic fibroblast." The present study attempts to explore the ALPase synthesized on HPLF in relation to 1,25(OH)2D3. These HPLF were obtained by the explantation method and then subcultured in D-MEM containing 2 mg FCSP/ml, 50 micrograms ascorbic acid/ml and penicillin/streptomycin after trypsinization. The HPLF were inoculated at a cell density of 1.25 x 10(4) cells/cm2 in culture wells. After 24hr, the HPLF were treated every two days for 7 days with 0.5-10nM 1,25 (OH)2D3. Then, ALPase activity, DNA and protein contents were assayed by the methods using p-nitrophenylphosphate, diaminobenzoic acid and Coomassie Brilliant Blue, respectively. Also, ALPase was prepared from the confluent HPLF incubated with 5 nM 1,25 (OH)2D3 for 12 days, and digested with and without trypsin. The crude ALPase which was solubilized with 10mM Tris-HCl, pH 7.4 containing 0.2 mM MgCl2 and 0.1% NP-40 was applied to 5-15% gradient SDS-PAGE and stained with beta-naphththylacid phosphate and First Blue BB salt in 60 mM borate buffer pH 9.7. The cell growth which was assayed by DNA contents and the incorporation of 3H-thymidine was decreased by 1,25(OH)2D3. On the other hand, ALPase activity was increased approximately 3.6 fold at 6 day by the addition of 5 nM 1,25(OH)2D3. From the separation of ALPase activity on SDS-PAGE, 110 K and 120-130 K ALPase were identified. The 110 K ALPase, which was not changed by 1,25(OH)2D3, was converted to 100K, releasing 10K peptide after trypsin treatment. This 110K ALPase might be tightly associated with cell membrane structure. The 120-130K ALPase was remarkably increased by 1,25(OH)2D3 on SDS-PAGE and completely digested with trypsin. The ALPase in the cultured HPLF might be located not only on the plasma membrane but also in the extracellular matrix. Therefore, 1,25(OH)2D3 may regulate the cell cycle and also the gene expression of ALPase of HPLF.
...
PMID:[Biochemical study of human periodontal ligament fibroblasts--1,25 (OH)2D3 dependent alkaline phosphatase]. 248 52

Cleavage of bacterial alkaline phosphatase by trypsin at the R-11, A-12 bond of both subunits results in changes in the structure and function of the enzyme as previously reported (Roberts, C. H., and Chlebowski, J. F. (1984) J. Biol. Chem. 259, 729-733; Roberts, C. H., and Chlebowski, J. F. (1985) J. Biol. Chem. 260, 7557-7561). A hybrid dimer has been formed by cleaving the R-11, A-12 bond of only one of the two subunits. This enzyme species has been purified and characterized to investigate subunit interactions of this hybrid dimeric enzyme species. Subunit interactions were observed using various methods to study functional and structural properties of the enzyme. In a kinetic study the T-2/A-12 hybrid enzyme was found to have a Vmax similar to the A-12 fully trypsin-modified enzyme. On exposure to EDTA the hybrid was found to lose activity at essentially the same rate as the A-12 enzyme presumably as a consequence of loss of metal ions required for function. On adding metal ions back to the apoenzyme form, activity of the hybrid was reconstituted to a degree similar to that of the native enzyme whereas the activity of the A-12 enzyme was reconstituted to a much lesser extent. The Tm of the hybrid measured by differential scanning calorimetry was closer to the value obtained for the A-12 enzyme than the T-2 enzyme but circular dichroic spectra indicated secondary structural features of the hybrid different from both symmetrical forms of the enzyme. These results provide evidence for strong subunit interactions in the alkaline phosphatase dimer.
...
PMID:A hybrid Escherichia coli alkaline phosphatase formed on proteolysis. 249 74

Subcutaneous implantation of demineralized bone matrix (DBM) from rat initiates a sequence of developmental events that results in endochondral bone formation. This investigation examined the modification of the osteoinductive potential of DBM during the initial stages of this developmental cascade. Diffusion chambers (DC), constructed with filters of known pore size, permitting or excluding cells from entering the chambers, and containing DBM were subcutaneously implanted into Long-Evans male rats for specific time periods (1-7 days). DC were recovered and the osteoinductive potential of the matrix from these chambers was then tested by subcutaneous implantation and assaying the resulting day 11 plaque tissue enzymatically for alkaline phosphatase activity, and histologically for evidence of chondrogenesis and osteogenesis. The possible modification of DBM by local systemic factors (enzymatic degradation) or contact by polymorphonuclear leukocytes (PMNs) was also investigated. We have concluded from this study that the osteoinductive potential of DBM has a half-life of 5-7 days following implantation and although the enzymes collagenase, elastase, and trypsin abolished this activity, pepsin significantly enhanced it. Culture of PMNs with matrix prior to its implantation appeared to have little effect. Furthermore, during the initial stages of matrix-induced endochondral bone formation, DBM serves as both the instructive inducer and permissive substratum required in this process.
...
PMID:In vivo analysis of the half-life of the osteoinductive potential of demineralized bone matrix using diffusion chambers. 250 25

The osteoblast phenotype is characterized by its ability to (a) synthesize a well defined mineralized collagenous matrix, (b) regulate the remodeling process by synthesizing local hormones (PGE2) and specific molecules (osteocalcin) and enzymes (alkaline phosphatase and collagenase), (c) respond to a variety of hormones (PTH, PGs, vitamin-D metabolites, steroids and growth factors), (d) respond to mechanical stimulation. Most of osteoblast culture systems meet many of the above qualifications though most fail to show the PTH effect on DNA synthesis, (c), and mechanical stimulation (d). Here we show that by using trypsin digestion and serum-containing low calcium medium (0.25 mM), all the above listed osteoblast phenotypic characteristics are demonstrated including their responsiveness to mechanical stimulation and the PTH effect on DNA synthesis.
...
PMID:Calvaria derived osteogenic cells: phenotypic expression in culture. 261 64

Bacteroides gingivalis was grown in continuous culture in the presence of chlorhexidine. Maximum specific growth rates and biomass levels initially increased but then decreased as the chlorhexidine level increased from 0 to 30 micrograms/ml. Total inhibition of growth occurred when the chlorhexidine concentration reached 60 micrograms/ml. The steady-state levels of cell-bound, extracellular vesicle and extracellular soluble enzymes, trypsin-like protease, alkaline phosphatase and N-acetyl-beta-glucosaminidase were measured. With increasing sub-lethal concentrations of chlorhexidine, levels of alkaline phosphatase increased noticeably in all three fractions of culture, whilst cell-bound and extracellular vesicle levels of N-acetyl-beta-glucosaminidase remained approximately constant. Extracellular soluble levels of alkaline phosphatase and N-acetyl-beta-glucosaminidase increased with increasing levels of chlorhexidine. The levels of trypsin-like protease decreased significantly in all fractions of the culture when cells were grown in the presence of chlorhexidine. Thus, chlorhexidine has a differential effect on the production of B. gingivalis hydrolytic enzymes.
...
PMID:The effects of chlorhexidine on the maximum specific growth rate, biomass and hydrolytic enzyme production of Bacteroides gingivalis grown in continuous culture. 261 91

Bacteroides gingivalis strain W50 was grown in batch and continuous culture on complex medium with haemin. In batch culture, cell-bound levels of trypsin-like protease (EC 3.4.21.4), alkaline phosphatase (EC 3.1.3.1) and N-acetyl-beta-glucosaminidase (EC 3.2.1.30) increased during the exponential phase of growth. These enzyme activities were also detected in extracellular vesicles and in extracellular soluble forms in the supernatant fluid, but in lower amounts per unit biomass compared to cell-bound levels. In continuous culture, at high relative growth rates (0.7-0.9 murel), the highest proportions of enzyme activities were cell-bound. In contrast, at low relative growth rates (0.1-0.2 murel), highest enzyme levels were detected in the extracellular vesicle fraction. Levels of extracellular soluble enzymes were always low compared to cell-bound or extracellular vesicle levels, but were highest at low relative growth rates. All three enzymes appeared to be relatively stable in their soluble forms. Vesicle production appeared to be associated with actively growing cells but was influenced by growth rate. The results are consistent with the hypothesis that cell-bound 'periplasmic' enzymes are encapsulated into vesicles which are subsequently released by the cells. Therefore, levels of total extracellular enzyme (extracellular vesicle plus extracellular soluble) may depend on the rate of vesicle formation superimposed on the rates of production of 'periplasmic' enzymes in the cell.
...
PMID:Production of cell-bound and vesicle-associated trypsin-like protease, alkaline phosphatase and N-acetyl-beta-glucosaminidase by Bacteroides gingivalis strain W50. 262 40


<< Previous 1 2 3 4 5 6 7 8 9 10 Next >>

---