Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P00750 (PLA)
 16,800 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Synthetic oligonucleotides were used to amplify phospholipase A(2) (PLA(2)) gene by RT-PCR from total RNA of snake Agkistrodon acutus venom gland. The PCR products were subcloned and positive clones were screened with acidic PLA(2) gene from Agkistrodon halys Pallas. Finally, four cDNAs of PLA(2) isoenzymes were isolated. Their complete sequences were determined by bidirectional sequencing and their amino acid sequences were deduced. They were designated as A.aAPLA(2)I A.aAPLA(2)II A.aBPLA(2) and A.aLys(49)-PLA(2) according to their isoelectric points calculated by computer and special structure characteristics respectively. The amino acid sequence of 1 10 residues of A.aAPLA(2)I deduced from the cDNA is identical to that of acidic PLA(2) which had been isolated from Agkistrodon acutus. A.aLys(49)-PLA(2) is unique because of the usual Asp(49) is replaced by Lys(49), which may lower its enzymatic activity. Their similarity scores were calculated and compared by computer. The successful cloning of these isoenzymes genes may provide more information for the study on structure-function relationship of PLA(2) family.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai) 1999
PMID:Cloning and Sequencing of Genes Encoding Phospholipase A(2) from Agkistrodon acutus. 1214 13

We have modeled three-dimensional structures of basic-acidic hybrid phospholipase A(2)-II and neutral phospholipase A(2) from venom of snake Agkistrodon halys Pallas, based on the known structures of basic and acidic phospholipase A(2)'s from the same source. We have compared these structures of phospholipase A(2)'s, explained the results of fluorescent spectrum study on the phospholipase A(2)'s and calculated the electrostatic potential maps on the catalytic active site. We suggest that the electrostatic potential around the catalytic active site of PLA(2) containing a calcium ion favors the binding of the PLA(2) to its substrate with negative charge.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai) 1999
PMID:Modeling and Analysis of Structures of Phospholipase A(2)'s from Venom of Agkistrodon haly Pallas. 1214 15

PLA(2)(EC3.1.1.4) of Agkistrodon blomhoffii Ussurensis of Changbai Mountain has been purified to homogeneity via four steps consisting of DEAE-Sephadex A-50, and three times Sephadex G-75 column chromatography in series. Using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and matrix-assisted laser desorption/ionization, time of flight mass spectrometry, the PLA(2) was found to have a molecular weight of (14.008+/-0.007) kD. The optimal pH of the PLA(2) is in the range from 8.0 to 9.0, the optimal temperature is 45 degrees. Polymer is found in solution by MALDI/TOF/MS.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai) 1999
PMID:Purification and Characterization of A Phospholipase A(2) from Agkistrodon blomhoffii Ussurensis of Changbai Mountains. 1214 27

To obtain a thromblytic agent with high efficacy and specifity, we have engineered a recombinant chimeric plasminogen activator SZ51Hu-scuPA, which was composed of a humanized monoclonal antibody (SZ51Hu) directed against activated human platelets and a single-chain urokinase (scuPA). The cDNA sequence encoding scuPA was fused through 5' end to 3' end of the CH(3) of SZ51Hu heavy-chain sequence in the expression vector alphalys30(alphalys30-SZ51VH/Hu-scuPA) by PCR. This construct was transfected into a murine myeloma line secreting SZ51Hu light chain with Lipofectin reagent and three lines which showed stable integration and high level expression with concentration of 5 mg/L in supernatant, were selected in the end. Purified SZ51Hu-scuPA migrated as a 160 kD band on non-reduced SDS/PAGE and had the same characteristic of binding to the activated platelets compared with its parent murine antibody molecule SZ51 by Western-blot analysis. The fibrinolytic activity of purified products was 39 000 IU/mg. Thus, by recombinant techniques, an expressed fusion protein was capable of specific binding to activated platelets and plasminogen activation. These observations laid a solid foundation for further study of targeting of thrombolysis in vitro and in vivo.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai) 1998
PMID:Construction and Expression of a Recombinant Antibody-targeted Plasminogen Activator Composed of a Humanized Monoclonal Antibody against Activated Human Platelets and a Single-chain Urokinase. 1216 96

An antineurotoxic factor has been purified from the serum of the snake B. multicinctus. The purified antineurotoxic factor shows a single band in PAGE and SDS-PAGE, running in the region of alpha(2)- or beta-globulin. This antineruotoxic factor is a glycoprotein and its molecular weight is (80+-5) kD. This antineurotoxic factor can partially neutralize the toxicity of beta-BuTx and possesses inhibition activity to the beta-BuTx PLA(2) enzyme.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai) 1998
PMID:Purification of an Antineurotoxic Factor from Serum of Snake Bungarus multicinctus. 1216 46

We used degenerate primers to amplify phospholipase A(2)(PLA(2)) gene by RT-PCR from venom total RNA of Agkistrodon halys Pallas, and screened with B-PLA(2) gene as probe. Finally we isolated three cDNAs containing A-PLA(2) and two other genes showing similar characteristic structure--Asn(49)-PLA(2) and BA-PLA(2). Their complete sequence was determined by bidirectional sequencing and their amino acid sequence was deduced. The amino acid sequence of A-PLA(2) deduced from the cDNA agreed with that determined by protein sequencing except for four residues; Asn(49 )-PLA(2) and B-PLA(2) are very alike (up to 95%), but the Asp(49)-PLA(2) of B-PLA(2) involved in the enzyme reaction is replaced by Asn(49) of Asn(49)-PLA(2). Therefore, it possibly affects the enzyme reaction of Asn(49)-PLA(2). The N-terminus sequence of BA-PLA(2) is highly homologous to B-PLA(2), but its C-terminus sequence is almost the same as A-PLA(2). The successful cloning of these isoenzyme genes not only discloses the structure diversity of PLA(2)(namely DNA modification and recombination), but also provides excellent material for the study of structure-function relationship in PLA(2).
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai) 1998
PMID:Research on the Diversity of PLA(2) Gene from Agkistrodon halys Pallas. 1217 5

HPLC was used to analyse the drug concentrations in blood. The 3p87 (practical pharmacokinetic program-version) was used to calculate the parameters of DHAQ-PLA-NP lyophilization injection and DHAQ injection through i.v. administration in rabbit blood. The concentration-time curves of the two kinds of preparations were in keeping with the three-compartment model. The drug concentrations were calculated according to the selecting pharmaceutical model and parameters. The calculated drug concentrations agreed well with the observed. The lyophilization injection sustained the eliminating process in vivo.
Hua Xi Yi Ke Da Xue Xue Bao 1999 Sep
PMID:[Study on the pharmacokinetics of the liver targeted mitoxantrone polylactic acid extended-release nanoparticles in rabbits]. 1221 3

A phospholipase A(2)(PLA(2)) was purified from the venom of the snake Trimeresurus stejnegeri Schmidt by Sephadex G-75 gel filtration, Mono Q ion exchange chromatography and Superose-12 gel filtration with FPLC and proved to be homogeneous as shown in SDS-PAGE and IEF. Its molecular weight is around 18 000 and isoelectric point is 4.7. Amino acid composition of PLA(2) was determined. Apart from hydrolyzing phosphatidylcholine, the enzyme has a potent inhibitory effect on platelet aggregation induced by ADP or collagen with half-inhibitory concentration of 10-15 &mgr;g/ml and 50-80 &mgr;g/ml respectively.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai) 1996
PMID:Purification and Characterization of Phospholipase A(2) from the Venom of Snake Trimeresurus stejnegeri Schmidt. 1223 3

A 54 kD calcium-binding protein was isolated form porcine platelet after calcium precipitation and chromatography. Its calcium-blinding ability was proved by the Arasenzo III binding analysis. In order to study the physiology role of calcium-binding protein in platelets, an experiment to study the effect of 54 kD calcium-binding protein on the platelet membrane-bound PLA(2) was designed. We isolated porcine platelet membrane by the two-phases system and developed a simple assay method for phospholipase A(2) by the colorimetric micro-determination of long-chain fatty acid released by the enzyme. The results indicated that in the presence of 54 kD CaBP the activity of porcine platelet membrane-bound PLA(2) was stimulated with lower Ca(2+) concentration (<1 mM), but inhibited when the Ca(2+) concentration was above 1 mM. The inhibiting effect of 54 kD CaBP was not decreased with the increase of substrate concentration.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai) 1996
PMID:The Effect of 54 kD Calcium-binding Protein on the Platelet Membrane-bound PLA(2). 1223 21


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