Uturu hwenyoka hune ma enzymes ano hydrolyze carboxyl ester bond.Iyo substrates ye hydrolysis ndeye phospholipids, acetylcholine uye inonhuhwirira acetate.Aya ma enzymes anosanganisira marudzi matatu: phospholipase, acetylcholinesterase uye anonhuwirira esterase.Arginine esterase muhuturu hwenyoka inogona zvakare kuita hydrolyze synthetic arginine kana lysine, asi inonyanya kuita hydrolyze maprotein peptide bond muzvisikwa, saka ndeyeprotease.Ma enzymes anokurukurwa pano anongoshanda pane ester substrates uye haagone kuita pane chero peptide bond.Pakati pema enzymes aya, mabasa ehupenyu eacetylcholinesterase uye phospholipase anonyanya kukosha uye akanyatsodzidza.Humwe huturu hwenyoka hune simba rakasimba rekuita esterase, rinogona hydrolyze p-nitrophenyl ethyl ester, a - kana P-naphthalene acetate uye indole ethyl ester.Izvo hazvisati zvazivikanwa kana chiitiko ichi chinogadzirwa neiyo enzyme yakazvimiririra kana inozivikanwa divi mhedzisiro yecarboxylesterase, tisiye kukosha kwayo kwebiological.Apo uturu hweAgkistrodon halys Japonicus hwakaitwa ne p-nitrophenyl ethyl ester uye indole ethyl ester, iyo hydrolysates ye p-nitrophenol uye indole phenol haina kuwanikwa;Pane zvinopesana, kana esters aya akaita necobra Zhoushan subspecies nyoka uturu uye Bungarus multicinctus nyoka uturu, ivo vachakurumidza hydrolyzed.Zvinozivikanwa kuti uturu hwemhungu hune simba recholinesterase chiitiko, chinogona kunge chiri chekuita hydrolysis yeaya ari pamusoro apa.Muchokwadi, Mclean et al.(1971) yakashuma kuti huturu hwenyoka huzhinji hwemhuri yeCobra hunogona hydrolyze indole ethyl ester, naphthalene ethyl ester uye butyl naphthalene ester.Huturu hwenyoka uhu hunobva: mhungu, mhungu nhema, mhungu nhema, mhungu yegoridhe, mhungu yeEgipita, king cobra, golden cobra mamba, black mamba newhite lipped mamba (D. aw achiri kuziva eastern rhombola rattlesnake
Uturu hwenyoka hunogona kuita hydrolyze methyl indole ethyl ester, inova iyo substrate yekuona chiitiko checholinesterase muserum, asi uturu hwenyoka uhu hahuratidzi chiitiko checholinesterase.Izvi zvinoratidza kuti kune esterase isingazivikanwe muhuturu hwemhungu, hwakasiyana necholinesterase.Kuti unzwisise chimiro cheiyo enzyme, rimwe basa rekuparadzanisa rinodiwa.
1, Phospholipase A2
(I) Muchidimbu
Phospholipase is enzyme inogona hydrolyze glyceryl phosphate.Kune marudzi mashanu e phospholipase muzvisikwa, anoti phospholipase A2 uye phospholipase.
A. , phospholipase B, phospholipase C uye phospholipase D. Uturu hwenyoka hunonyanya kuve nephospholipase A2 (PLA2), uturu hwenyoka hushoma hune phospholipase B, uye mamwe maphospholipases anonyanya kuwanikwa munyama yemhuka uye mabhakitiriya.Mufananidzo 3-11-4 inoratidza nzvimbo yekuita yeiyi phospholipases pane substrate hydrolysis.
Pakati pephospholipases, PLA2 yakadzidzwa zvakanyanya.Inogona kunge iri iyo enzyme yakanyanya kudzidzwa muhuturu hwenyoka.Yayo substrate ndiyo ester bond panzvimbo yechipiri yeSn-3-glycerophosphate.Enzyme iyi inowanikwa zvakanyanya muhuturu hwenyoka, uturu hwenyuchi, huturu hwescorpion uye matishu emhuka, uye PLA2 yakawanda muhuturu hwenyoka hwemhuri.Nekuti iyi enzyme inotyora masero matsvuku eropa uye inokonzera hemolysis, inonziwo "hemolysin".Vamwe vanhu vanodana zvakare PLA2 hemolytic lecithinase.
Ludeeke akatanga kuona kuti uturu hwenyoka hunogona kuburitsa musanganiswa wehemolytic nekuita pane lecithin kuburikidza nema enzymes.Gare gare, Delezenne et al.yakaratidza kuti kana uturu hwemhungu huchibata paserum yebhiza kana yolk, inoumba chinhu chehemolytic.Iko zvino yava kuzivikanwa kuti PLA2 inogona kuita zvakananga pane phospholipids ye erythrocyte membrane, ichiparadza chimiro che erythrocyte membrane uye kukonzera zvakananga hemolysis;Inogona zvakare kuita paserum kana yakawedzera lecithin kugadzira hemolytic lecithin, inoshanda pamasero matsvuku eropa kugadzira isina kunanga hemolysis.Kunyange zvazvo PLA2 yakawanda mumhuri ina dzehuturu hwenyoka, zviri mukati ma enzymes muhuturu hwenyoka hwakasiyana zvakasiyana.Rattlesnake (C
Uturu hwenyoka hwaingoratidza husina simba hwePLA2 kuita.Tafura 3-11-11 inoratidza kuenzanisa kwePLA2 chiitiko chegumi huturu hukuru hwenyoka dzine uturu muChina.
Tafura 3-11-11 Kuenzanisa kwe phospholipase VIII zviitiko zve 10 huturu hwenyoka muChina.
Uturu hwenyoka
Kusunungurwa kwemafuta
Aliphatic acid,
Cjumol/mg)
Hemolytic chiitiko CHU50/^ g * ml)
Uturu hwenyoka
Bvisa mafuta acids
(^raol/mg)
Hemolytic chiitiko "(HU50/ftg * 1111)
Najanaja atra
9. 62
gumi neimwe
Micracephal ophis
shanu point one zero
kalyspallas
8. 68
zviuru zviviri nemazana masere
gracilis
V, acutus
7. 56
* * #
Ofiophagus Hana
nhatu pfungwa sere mbiri
zana namakumi mana
Bnugarus fasctus
7,56
mazana maviri nemakumi masere
B. multicinctus
imwe pfungwa pfumbamwe nhanhatu
mazana maviri nemakumi masere
Viper uye russelli
nomwe point zero three
T, mucrosquamatus
imwe pfungwa sere shanu
Siamensis
T. stejnegeri
0. 97
(2) Kuparadzaniswa uye kucheneswa
Izvo zviri muPLA2 muhuturu hwenyoka yakakura, uye yakagadzikana pakupisa, asidhi, alkali uye denaturant, kuitira kuti zvive nyore kuchenesa nekuparadzanisa PLA2.Nzira yakajairika ndeyekutanga kuita kusefa gel pahuturu husina kuchena, wozoita ion exchange chromatography, uye nhanho inotevera inogona kudzokororwa.Izvo zvinofanirwa kucherechedzwa kuti kuomesa-kuomeswa kwePLA2 mushure meiyo ion-exchange chromatography haifanire kukonzera kuunganidzwa, nekuti kuomesa-kuomesa kunowanzo kuwedzera simba reionic muhurongwa, icho chinhu chakakosha chinokonzera kuunganidzwa kwePLA2.Pamusoro penzira dziri pamusoro apa, nzira dzinotevera dzakagamuchirwa: ① Wells et al.② Iyo substrate analogue yePLA2 yakashandiswa se ligand yekubatana chromatography.Iyi ligand inogona kusunga kuPLA2 muhuturu hwenyoka neCa2+.EDTA inonyanya kushandiswa seyakajeka.Mushure mokunge Ca2 + yabviswa, hukama huri pakati pePLA2 uye ligand hunoderera, uye hunogona kuparadzaniswa kubva kune ligand.Vamwe vanoshandisa 30% organic solution kana 6mol/L urea sevanoziva.③ Hydrophobic chromatography yakaitwa nePheiiylSephar0SeCL-4B kubvisa trace PLA2 mu cardiotoxin.④ Anti PLA2 antibody yakashandiswa se ligand kuita affinity chromatography paPLA2.
Kusvika ikozvino, nhamba huru yehuturu hwenyoka PLAZ yakacheneswa.Tu et al.(1977) yakanyorwa PLA2 yakacheneswa kubva muhuturu hwenyoka pamberi pe 1975. Mumakore achangopfuura, nhamba huru yezvinyorwa pamusoro pekuparadzanisa uye kucheneswa kwePLA2 zvakataurwa gore negore.Pano, isu tinotarisa pakuparadzaniswa uye kucheneswa kwePLA nenyanzvi dzeChinese.
Chen Yuancong et al.(1981) yakapatsanura matatu emhando dzePLA2 kubva muhuturu hweAgkistrodon halys Pallas muZhejiang, iyo inogona kupatsanurwa kuita acidic, kwayakarerekera uye alkaline PLA2 zvinoenderana neisoelectric point.Zvinoenderana nehupfu hwayo, kusarerekera kwePLA2 kune muchetura zvakanyanya, izvo zvakaonekwa se presynaptic neurotoxin Agkistrodotoxin.Alkaline PLA2 haina muchetura, uye acidic PLA2 inenge isina chepfu.Wu Xiangfu et al.(1984) akaenzanisa hunhu hwemaPLA2 matatu, kusanganisira huremu hwemamorekuru, amino acid kuumbwa, N-terminal, isoelectric point, thermal stability, enzyme activity, toxicity uye hemolytic basa.Mhedzisiro yacho yakaratidza kuti vaive nehuremu hwakafanana hwemamorekuru uye kugadzikana kwekupisa, asi vaive nekusiyana kwakakura mune zvimwe zvinhu.Muchikamu chebasa re enzyme, acid enzyme basa raive rakakwirira kupfuura alkaline enzyme basa;Iyo hemolytic mhedzisiro yealkaline enzyme pamakonzo masero matsvuku eropa ndiyo yaive yakasimba, ichiteverwa neyakarerekera enzyme, uye asidhi enzyme isingaite hemolyzed.Nokudaro, zvinofungidzirwa kuti hemolytic effect yePLAZ yakabatana nemhosva yePLA2 molecule.Zhang Jingkang et al.(1981) vakagadzira Agkistrodotoxin makristasi.Tu Guangliang et al.(1983) yakashuma kuti PLA ine chepfu ine isoelectric point ye 7. 6 yakaparadzaniswa uye yakacheneswa kubva muhuturu hweVipera rotundus kubva kuFujian, uye zvayo zvepanyama uye zvemakemikari, amino acid kuumbwa uye kutevedzana kwe22 amino acid zvakasara paN. -terminal yakatemwa.Li Yuesheng et al.(1985) akazviparadzanisa uye akachenesa imwe PLA2 kubva muhuturu hweViper rotundus muFujian.Iyo subunit yePLA2 * ndeye 13 800, isoelectric point i10.4, uye chaiyo chiitiko i35/xnioI/miri mg. Ne lecithin se substrate, iyo yakakwana pH ye enzyme i8.0 uye tembiricha yakakwana i65 ° C. LD5 inobaiwa mutsinga mumakonzo.Iyo 0.5 ± 0.12mg/kg.Iyi enzyme ine pachena anticoagulant uye hemolytic mhedzisiro.Muchetura wePLA2 morekuru une 123 masara emarudzi gumi nemasere emaamino acids.Iyo molecule yakapfuma mu cysteine (14), aspartic acid (14) uye glycine (12), asi ine methionine imwe chete, uye N-terminal yayo inosara serine.Kuenzaniswa nePLA2 yakaparadzaniswa neTuguang, huremu hwema molecular uye nhamba yeamino acid residues yemasoenzymes maviri akafanana zvikuru, uye amino acid yakagadzirwa yakafanana zvikuru, asi nhamba yeaspartic acid uye proline residues yakasiyana.Guangxi king cobra snake venom ine hupfumi PLA2.Shu Yuyan et al.(1989) yakatsaura PLA2 kubva muhuturu, iyo ine chiitiko chakanyanya 3.6 yakakwira kupfuura yehuturu hwepakutanga, molecular huremu hwe13000, kuumbwa kwe122 amino acid zvakasara, isoelectric point ye8.9, uye yakanaka kugadzikana kwekupisa.Kubva pakuona maelectronic microscope mabatiro eBasic PLA2 pamasero matsvuku eropa, zvinoonekwa kuti zvine mhedzisiro pamasero matsvuku eropa remunhu, asi hazvina zvazvinoita pamasero matsvuku eropa.Iyi PLA2 ine mhedzisiro yekunonoka kwemagetsi pakumhanya kwemasero matsvuku eropa muvanhu, mbudzi, tsuro uye guinea pigs.Chen et al.Iyi enzyme inogona kurambidza kuunganidzwa kweplatelet kunokonzerwa neADP, collagen uye sodium arachidonic acid.Kana PLA2 yakasungwa iri 10/xg/ml~lOOjug/ml, kuunganidzwa kweplatelet kunorambidzwa zvachose.Kana maplatelet akashambidzwa akashandiswa semidziyo, PLA2 yaisagona kurambidza kuunganidzwa pakusangana kwe20Mg/ml.Aspirin inhibitor ye cyclooxygenase, iyo inogona kudzivisa kuitika kwePLA2 pamaplatelet.PLA2 inogona kurambidza kuunganidzwa kweplatelet ne hydrolyzing arachidonic acid kugadzira thromboxane A2.Mhinduro yePLA2 inogadzirwa neAgkistrodon halys Pallas uturu muZhejiang Province yakaongororwa nenzira yedenderedzwa dichroism, fluorescence uye UV kutorwa.Mhedzisiro yekuedza yakaratidza kuti cheni huru yekuumbwa kweiyo enzyme yaive yakafanana neyemhando imwechete ye enzyme kubva kune mamwe marudzi uye genera, iyo skeleton conformation yaive neyakanaka kupisa kupisa, uye shanduko yemaitiro munzvimbo yeasidhi yakadzoserwa.Iko kusanganiswa kwe activator Ca2 + uye enzyme haikanganisi nharaunda yemasara e tryptophan, nepo inhibitor Zn2 + ichiita zvinopesana.Iyo nzira iyo pH kukosha kwemhinduro inokanganisa kuita kwe enzyme yakasiyana nemareagents ari pamusoro.
Mukuita kwePLA2 kucheneswa kwehuturu hwenyoka, chinhu chiri pachena ndechekuti huturu hwenyoka hune maviri kana anopfuura ePLA2 elution peaks.Ichi chiitiko chinogona kutsanangurwa sezvizvi: ① nekuda kwekuvapo kwemaisozymes;② Imwe mhando yePLA2 inoiswa polymer mumhando dzakasiyana dzePLA2 misanganiswa ine uremu hwakasiyana-siyana hwemamorekuru, mazhinji acho ari muchikamu che9 000 ~ 40 000;③ Kusanganiswa kwePLA2 uye zvimwe zvikamu zvehuturu hwenyoka zvinokanganisa PLA2;④ Nekuti iyo amide bond muPLA2 ine hydrolyzed, mutero unochinja.① Uye ② zvakajairika, zvichingosara zvishoma, senge PLA2 muCrWa/w uturu hwenyoka.
Pane mamiriro maviri: ① uye ②.Mamiriro echitatu akawanikwa muPLA2 muhuturu hwenyoka dzinotevera: Oxyranus scutellatus, Parademansia microlepidota, Bothrops a ^> er, nyoka yePalestine, nyoka yejecha, uye inotyisa rattlesnake km.
Mhedzisiro yenyaya ④ inoita kuti kumhanya kwePLA2 kuchinjike panguva ye electrophoresis, asi iyo amino acid inoumbwa haichinji.Peptides inogona kutyorwa ne hydrolysis, asi kazhinji ichiri kusungwa pamwe chete ne disulfide bond.Huturu hweeastern pit rattlesnake ine mafomu maviri ePLA2, anonzi type a uye type p PLA2 zvakateerana.Musiyano uripo pakati pemhando mbiri idzi dzePLA2 inongova imwe amino acid, kureva, glutamine mune imwe PLA2 molecule inotsiviwa ne glutamic acid mune imwe PLA2 molecule.Kunyangwe chikonzero chaicho chemutsauko uyu chisina kujeka, zvinotendwa kuti zvine hukama nekubviswa kwePLA2.Kana PLA2 muPalestine huturu hwenyoka ikachengetwa ichidziya nehuturu husina kuchena, mapoka ekupedzisira mumamorekuru ayo eenzayimu anozowanda kupfuura kare.Kubva kuC PLA2 yakaparadzaniswa nehuturu hwenyoka ine N-terminal mbiri dzakasiyana, uye molecular uremu i30000. Chiitiko ichi chinogona kukonzerwa neasymmetric dimer yePLA2, yakafanana nesymmetric dimer inoumbwa nePLA2 muhuturu hwe eastern diamondback rattlesnake. uye western diamondback rattlesnake.Mhungu yekuAsia inoumbwa nemhando dzakawanda, dzimwe dzacho dzisina kunyatsojeka mukuronga.Semuenzaniso, yaimbonzi Cobra Outer Caspian subspecies yava kuzivikanwa
Inofanira kuverengerwa kune Outer Caspian Sea Cobra.Sezvo kune akawanda masubspecies uye akasanganiswa pamwechete, kuumbwa kwehuturu hwenyoka kunosiyana zvakanyanya nekuda kwenzvimbo dzakasiyana, uye zvirimo zvePLA2 isozymes zvakare zvakakwirira.Somuenzaniso, cobra uturu
Anosvika mapfumbamwe emhando dzePLA2 isozymes dze r ^ ll marudzi akawanikwa mukati, uye 7 marudzi emaPLA2 isozymes akawanikwa muhuturu hwecobra subspecies Caspian.Durkin et al.(1981) akadzidza zvirimo zvePLA2 uye huwandu hwemasozymes muhuturu hwenyoka hwakasiyana, kusanganisira huturu hwemhungu gumi nemasere, huturu hwemamba 3, huturu hwenyoka 5, huturu hwenyoka 16 uye huturu hwenyoka dzemugungwa 3.Kazhinji, chiitiko chePLA2 chehuturu hwemhungu chakakwira, chine ma isozymes akawanda.Chiitiko chePLA2 uye isozymes yehuturu hwenyoka zviri pakati nepakati.Chiitiko chePLA2 chehuturu hwemamba uye huturu hwenyoka hwakaderera kana kuti hapana chiitiko chePLA2.Chiitiko chePLA2 chehuturu hwenyoka yegungwa zvakare chakaderera.
Mumakore achangopfuura, hazvina kutaurwa kuti PLA2 muhuturu hwenyoka huripo muchimiro chezvinoita dimer, senge eastern rhombophora rattlesnake (C. snake venom ine type a uye type P PLA2, ose ari maviri anoumbwa nezvikamu zviviri zvakafanana. , uye chete dimerase ine
Chiitiko.Shen et al.zvakare akakurudzira kuti chete dimer yePLA2 yehuturu hwenyoka ndiyo inoshanda fomu ye enzyme.Kuongorora kwemamiriro ekunze kunoratidzawo kuti PLA2 yeWestern diamondback rattlesnake iripo muchimiro che dimer.Piscivorous compound
Kune maviri akasiyana PLA ^ Ei uye E2 yehuturu hwenyoka, umo 仏 iripo muchimiro che dimer, iyo dimer inoshanda, uye yayo dissociated monoma haishande.Lu Yinghua et al.(1980) akawedzera kudzidza maitiro emuviri nemakemikari uye reaction kinetics yaE. Jayanthi et al.(1989) yakatsaura PLA2 yakakosha (VRVPL-V) kubva muhuturu hwenyoka.Huremu hwemamorekuru hwemonomer PLA2 i10000, ine zvinouraya, anticoagulant uye edema mhedzisiro.Iyo enzyme inogona polymerize mapolymer ane akasiyana mamolecular uremu pasi pemamiriro ePH 4.8, uye dhigirii repolymerization uye huremu hwemamorekuru hwemapolymers hunowedzera nekuwedzera kwekushisa.Huremu hwepolymer hunogadzirwa pa96 ° C i53 100, uye chiitiko chePLA2 chepolymer iyi chinowedzera nembiri.
Nguva yekutumira: Nov-18-2022