Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Kringles are protein modules found within a wide variety of fibrinolytic and coagulation-related proteins that show binding affinity for lysine, lysine analogs and for fibrin. We report here the crystal structures of apolipoprotein(a) kringle IV37 (apo(a) K4(37)) in its free state and in separate complexes with two omega-amino acids, 6-aminohexanoic acid (6AHA) and p-aminomethylbenzoic acid (PAMBA). The structures of the unliganded form and of both complexes have been determined and refined by restrained least-squares methods to about 2.0 angstrom. The overall kringle architecture is essentially identical with that determined in other kringles but it shows some small significant structural changes in the lysine binding site. Ther is virtually no difference in conformation between the unliganded and complexed forms, suggesting that apo(a) K4(37) does not undergo any conformational rearrangement upon binding. The 6AHA molecule binds to apo(a) K4(37) in a completely different way from that observed with the kringle 4 of plasminogen (PGK4). Its amino group makes an ion pair interaction with the two aspartate residues (Asp55/Asp57) of the anionic center and its carboxylate group faces out into the solvent making water-mediated contacts with the protein. The mode of binding of PAMBA resembles more that decribed for 6AHA when bound to PGK4. The PAMBA molecule is bound by ion pair interactions with the two aspartate residues (Asp55/Asp57) and with Arg71 from the cationic center and by van der Waals contacts. The relative importance of the cationic center from kringles for binding zwitterionic ligands is discussed.
J Mol Biol 1996 Mar 08
PMID:Crystal structures of apolipoprotein(a) kringle IV37 free and complexed with 6-aminohexanoic acid and with p-aminomethylbenzoic acid: existence of novel and expected binding modes. 864 95

Binding of urokinase-type plasminogen activator (uPA) to a specific receptor (uPAR) on human lung fibroblasts enables it to regulate cellular proteolysis and remodeling of the extracellular matrix. Binding studies with radiolabeled uPA indicated that both normal and fibrotic lung fibroblasts express the receptor, but cells from fibrotic tissues bound significantly more uPA (P < 0.001). Phorbol myristate acetate, lipopolysaccharide, transforming growth factor-beta (TGF-beta), and tumor necrosis factor-alpha (TNF-alpha) increased uPA binding and plasminogen activation at the cell surface, with a greater maximal effect on fibrotic than on normal fibroblasts. Excess unlabeled uPA, specific antibody, or antisense oligonucleotides inhibited uPA binding. Ribonuclease (RNase) protection assays showed higher levels of uPAR messenger ribonuleic acid (mRNA) in each of the five fibrotic cell lines than in normal fibroblasts. uPA was mitogenic for normal as well as fibrotic fibroblasts, indicating that receptor binding concurrently localizes cellular proteolytic activity and stimulates mitogenesis. Morphometry and immunohistochemical analysis showed that uPAR, as well as uPA, was increased in fibroblasts in fibrotic lung tissue. Increased expression of uPAR by fibrotic lung fibroblasts and enhanced urokinase binding induced by proinflammatory cytokines suggest a novel mechanism by which fibroblast-mediated matrix remodeling and proliferation may be regulated in interstitial lung diseases.
Am J Respir Cell Mol Biol 1996 Jul
PMID:Differential expression of the urokinase receptor in fibroblasts from normal and fibrotic human lungs. 867 25

Artificially induced rat decidual tissue expresses plasminogen activator inhibitor (PAI). This PAI, isolated and purified employing chromatographic techniques, is a low molecular weight protein unlike the known PAIs. The final purified preparation resolves into a single band following SDS-PAGE and has an approximate molecular weight of 29 kDa. The properties studied include specificity for urokinase-type (uPA) and tissue-type (tPA) plasminogen activators, binding to conA and heparin, inhibition of thrombin, plasmin and trypsin. Decidual PAI is immunogenic in rabbit and a monospecific antiserum raised against the decidual inhibitor cross reacts with an extract of human placenta.
Biochem Mol Biol Int 1995 Dec
PMID:A molecular variant of plasminogen activator inhibitor of rat decidual tissue. 874 33

The conversion of plasminogen to plasmin catalyzed by tissue-type plasminogen activator is accelerated in the presence of fibrin, leading to an increased dissolution of fibrin clots. This rate enhancement is mimicked by a dodecapeptide segment of the fibrin molecule containing the sequence A alpha 149-160 (RLEVDIDIKIRS). At low concentrations of the dodecapeptide, the potentiation increases with concentration, but at high concentrations, the stimulation effect diminishes, giving rise to a bell-shaped curve. The maximum rate enhancement of about 10 fold is achieved at a concentration of 85 micrograms/mL. This concentration dependent phenomenon is also observed for two synthetic peptide analogues, GLEVDIDIKIRS and RGGGGGGGKIRS, although the acceleration potential is less. These results indicate that the N-terminal amino acids are not critical for the rate enhancement. The bell-shaped activity-concentration curve suggests that the dodecapeptide may bind to both plasminogen and t-PA. This speculation is further supported by the modification of the potentiator. When the A alpha 149-160 dodecapeptide is pretreated with trypsin or phenyl glyoxal, the potentiation activity is eliminated. We speculate that the acceleration of the plasminogen-to-plasmin reaction catalyzed by t-PA is achieved through the action of the stimulator to bring the enzyme and its substrate together as a bi-dentate cross-linker. This effect increases the apparent concentration of the substrate at the enzyme active site, and is reflected as a decrease in Michaelis-Menten constant.
Biochem Mol Biol Int 1995 Dec
PMID:Mechanism of potentiation of tissue-type plasminogen activator. 874 37

Surface-associated plasmin(ogen) may contribute to the invasive properties of various cells. Analysis of plasmin(ogen)-binding surface proteins is therefore of interest. The N-terminal variable regions of M-like (ML) proteins from five different group A streptococcal serotypes (33, 41, 52, 53 and 56) exhibiting the plasminogen-binding phenotype were cloned and expressed in Escherichia coli. The recombinant proteins all bound plasminogen with high affinity. The binding involved the kringle domains of plasminogen and was blocked by a lysine analogue, 6-aminohexanoic acid, indicating that lysine residues in the M-like proteins participate in the interaction. Sequence analysis revealed that the proteins contain common 13-16-amino-acid tandem repeats, each with a single central lysine residue. Experiments with fusion proteins and a 30-amino-acid synthetic peptide demonstrated that these repeats harbour the major plasminogen-binding site in the ML53 protein, as well as a binding site for the tissue-type plasminogen activator. Replacement of the lysine in the first repeat with alanine reduced the plasminogen-binding capacity of the ML53 protein by 80%. The results precisely localize the binding domain in a plasminogen surface receptor, thereby providing a unique ligand for the analysis of interactions between kringles and proteins with internal kringle-binding determinants.
Mol Microbiol 1995 Nov
PMID:Identification of a plasminogen-binding motif in PAM, a bacterial surface protein. 874 39

Pathology of the prostate gland in rats and humans is associated with aging. Our objective was to examine the effects of aging on the activities of plasminogen activators and metalloproteases in the prostatic complex of rats. Plasminogen activator activities (very low in the anterior, lateral, and dorsal lobes, in contrast to higher activities in the ventral lobe of 4-month-old adult rats) increased with aging in the dorsal and anterior prostate lobes of 31-month-old rats; these activities also increased in the dorsal and lateral lobes upon castration of 18-month-old rats. The plasminogen activator activities in the ventral lobe did not increase with aging to 18 months but did increase 3-5-fold after castration of either young or old rats. Metalloprotease activities of 70 and 76 kDa were observed in the anterior and lateral lobes of 4 month untreated adult rats, whereas the dorsal lobe showed MP of 70 and 92 kDa. Castration of young adult rats increased activities of all three molecular forms of metalloprotease in these three lobes. Increased expression of metalloprotease activities was also found with aging to 31 months in the anterior, lateral, and dorsal lobes. However, changes in metalloprotease activities associated with age were most striking in the lateral lobe and included activities of 52, 55, 81, 93, 113, and 117 kDa at 18 months of age. Castration for 30 days at this age resulted in a decline in the 52, 55, 113, and 117 kDa activities and an increase in activities of the 70, 81, and 93 kDa forms. These latter metalloprotease activities were also increased in the dorsal lobe after castration. Our results suggest that some metalloprotease activities increased in the dorsal lobe after castration. Our results suggest that some metalloprotease activities increased in the lateral lobe with age possibly result from an increased accumulation of secretory proteins (i.e., 52, 55, 113, and 117 kDa), whereas the 70, 81, and 93 kDa metalloprotease activities may be related to possible prostatitis and/or involved in changes in tissue organization. The increased expression of metalloprotease activities in the lateral and dorsal prostate lobes with aging, and castration upon aging, may be indicative of altered hormonal regulation of these proteases in these lobes.
Cell Mol Biol Res 1995
PMID:Effects of aging and castration on plasminogen activator and metalloprotease activities in the rat prostate complex. 877 40

Plasminogen activator inhibitor 1 (PAI-1) is an important regulator of plasminogen activation, which inhibits both tissue-type plasminogen activator (tPA) and urokinase-type plasminogen activator (uPA). The DNA sequence encoding mature PAI-1 protein was inserted into an inducible expression vector. This gene was highly expressed to produce a soluble active protein in E. coli cells. The amount of the recombinant protein was up to 20% of total cellular protein. By efficient purification with a yield of about 15-20%, the recombinant protein could be purified to homogeneity with its specific activity up to 6.1 x 10(4) (uPA) IU/mg. Its inhibitory activity declined during incubation at 37 degrees C with a half life of about 2 hr.
Biochem Mol Biol Int 1996 May
PMID:High-level expression of active human plasminogen activator inhibitor type 1 (PAI-1) in E. coli. 879 49

Two low molecular weight urokinase derivatives were obtained by covalent coupling of a synthetic Gly-Pro-Arg-Pro tetrapeptide to peptide A of low molecular weight urokinase and exchanging the native peptide A of low molecular weight urokinase with Gly-Pro-Arg-Pro-peptide A to obtain derivative I or direct conjugation of Gly-Pro-Arg-Pro tetrapeptide to low molecular weight urokinase to obtain derivative II. In caseinolytic assay, fibrin can stimulate the two derivatives to activate plasminogen. But the two derivatives showed different kinetic behaviors. The derivative I displayed immediate onset of lysis and derivative II displayed a lag phase.
Biochem Mol Biol Int 1996 Jun
PMID:Chemical conjugation of Gly-Pro-Arg-Pro. Tetrapeptide to low molecular weight urokinase. 882 96

Human heart matrix metalloproteinases (MMP) are present in the latent form and activated in the failing heart. To examine whether the MMP activation was due to gene and/or post-translational modification, we analysed tissue from 10 explanted hearts due to coronary heart disease (CHD) and five normal left atrial tissue from donor hearts. Based on in situ immunolabeling MMP-1, tissue inhibitor of metalloproteinase (TIMP-1) and collagen were co-localized in the interstitial tissue. Based on sandwich ELISA, TIMP-1 and MMP-1 levels were 37 +/- 8 ng/mg and 9 +/- 2 ng/mg in normal tissue (P < 0.01) and 12 +/- 5 ng/mg and 75 +/- 11 ng/mg in the infarcted tissue (P < 0.01), respectively. These levels suggest repression of TIMP-1 during myocardial infarction. Northern blot analysis indicated that the mRNAs for both MMP-1 and TIMP-1 were increased three-to four-fold in the infarcted tissue as compared to the normal tissue, suggesting upregulation of MMP and TIMP gene transcription following infarction. Based on in situ tissue overlay zymography, the generalized activation of MMP was observed in the interstitium of the infarcted heart. Zymographic and immunoblot analysis demonstrated the presence of one band at 66 kDa (MMP-2) in the normal tissue and several bands at 92 (MMP-9), 66 (MMP-2) and 54 kDa (MMP-1) in the infarcted heart. Incubation of the zymographic gel with metal chelator (phenanthroline) abolished bands at 92 kDa and 54 kDa but phenanthroline did not abolish the lytic band at 66 kDa. The 66 kDa band was completely abolished in the presence of phenanthroline and phenyl methyl sulfonyl fluoride (PMSF). 2D-zymographic analysis suggested that the lytic band at 66 kDa was a mixture of two neutral proteinases with different isoelectric point. Plasminogen/gelatin zymographic analysis of infarcted tissue extract indicated that the band at 66 kDa was plasmin generated due to increased expression of tissue plasminogen activator (tPA) activity. In relation to increased expression of gelatinase in the infarcted tissue, our data suggest that gelatinase B (92 kDa) is induced in diseased heart. The results suggest that tPA converts plasminogen to plasmin which, in turn, activates MMPs and inactivates TIMP-1 post-translationally following ischemic cardiomyopathy.
J Mol Cell Cardiol 1996 Jul
PMID:Post-transcriptional regulation of extracellular matrix metalloproteinase in human heart end-stage failure secondary to ischemic cardiomyopathy. 884 29

A Gly-Pro-Arg-Pro tetrapeptide, homologous to amino-terminal segment of the human fibrin alpha chain after the release of the fibrinopeptide A, was covalently coupled to peptide A of low molecular weight urokinase. The resulting derivative gained increased affinity for fibrin. In caseinolytic assay, fibrin can stimulate the derivative to activate plasminogen. The derivative had two-fold greater fibrinolytic potency than native low molecular weight urokinase and its affinity for fibrin clot was 3.9-fold higher than that of low molecular weight urokinase.
Biochem Mol Biol Int 1996 Jul
PMID:A low molecular weight urokinase derivative with enhanced fibrin affinity. 884 49


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