Ubumara bwinzoka burimo enzymes hydrolyze carboxyl ester ihuza.Substrates ya hydrolysis ni fosifolipide, acetylcholine na acetate ya aromatic.Iyi misemburo irimo ubwoko butatu: fosifolipase, acetylcholinesterase na esterase ya aromatic.Arginine esterase muburozi bwinzoka irashobora kandi hydrolyze synthique arginine cyangwa lysine, ariko cyane cyane hydrolyzes protein peptide ihuza kamere, kubwibyo rero ni protease.Imisemburo yavuzwe hano ikora gusa kuri ester substrates kandi ntishobora gukora kuri peptide iyo ari yo yose.Muri iyi misemburo, imikorere yibinyabuzima ya acetylcholinesterase na fosifolipase ni ngombwa kandi yarizwe neza.Ubumara bumwe bwinzoka bufite ibikorwa bya esterase ikomeye ya aromatic, ishobora hydrolyze p-nitrophenyl Ethyl ester, a - cyangwa P-naphthalene acetate na estole estyl ester.Kugeza ubu ntiharamenyekana niba iki gikorwa cyakozwe na enzyme yigenga cyangwa ingaruka zizwi za carboxylesterase, tutibagiwe nubusobanuro bwibinyabuzima.Igihe uburozi bwa Agkistrodon halys Japonicus bwakiriwe na p-nitrophenyl etyl ester na indole etyl ester, hydrolysate ya p-nitropenol na fenol indole ntibyabonetse;Ibinyuranye nibyo, niba izo esters zifata hamwe nubumara bwinzoka ya cobra Zhoushan hamwe nuburozi bwinzoka ya Bungarus multicinctus, bizahita byoroha.Birazwi ko uburozi bwa cobra bufite ibikorwa bya cholinesterase bikomeye, bishobora kuba biterwa na hydrolysis ya substrate yavuzwe haruguru.Mubyukuri, Mclean n'abandi..Ubu bumara bwinzoka buturuka: cobra, cobra yijosi ryumukara, cobra yumukara wijimye, cobra yizahabu, cobra yo muri Egiputa, umwami cobra, zahabu cobra mamba, mamba yumukara na mamba yera yuzuye (D. aw iracyazi inzoka ya rhombola yuburasirazuba
Ubumara bwinzoka burashobora hydrolyze methyl indole ethyl ester, aribwo buryo bwo kumenya ibikorwa bya cholinesterase muri serumu, ariko ubu bumara bwinzoka ntabwo bugaragaza ibikorwa bya cholinesterase.Ibi birerekana ko hari esterase itazwi muburozi bwa cobra, butandukanye na cholinesterase.Kugira ngo wumve imiterere yiyi misemburo, hakenewe indi mirimo yo gutandukana.
1 、 Fosifolipase A2
(I) Incamake
Fosifolipase ni enzyme ishobora hydrolyze glyceryl fosifate.Hariho ubwoko 5 bwa fosifolipase muri kamere, aribyo fosifolipase A2 na fosifolipase
A..Igishushanyo cya 3-11-4 cyerekana urubuga rwibikorwa bya fosifolipase kuri hydrolysis ya substrate.
Muri fosifolipase, PLA2 yizwe byinshi.Irashobora kuba enzyme yize cyane muburozi bwinzoka.Substrate yayo ni ester ihuza kumwanya wa kabiri wa Sn-3-glycerophosphate.Iyi misemburo iboneka cyane muburozi bwinzoka, ubumara bwinzuki, ubumara bwa sikorupiyo nuduce twinyamanswa, kandi PLA2 ni nyinshi muburozi bune bwimiryango.Kubera ko iyi misemburo imena selile zitukura kandi igatera hemolysis, yitwa kandi "hemolysine".Abantu bamwe bita PLA2 hemolytic lecithinase.
Ludeeke yabanje kubona ko ubumara bwinzoka bushobora kubyara hemolytike ikora kuri lecithine ikoresheje enzymes.Nyuma, Delezenne n'abandi.yerekanye ko iyo ubumara bwa cobra bukora kuri serumu yamafarasi cyangwa umuhondo, bikora ibintu bya hemolytike.Ubu birazwi ko PLA2 ishobora gukora muburyo butaziguye kuri fosifolipide ya erythrocyte membrane, ikangiza imiterere ya membrane ya erythrocyte kandi igatera hemolysis itaziguye;Irashobora kandi gukora kuri serumu cyangwa kongeramo lecithine kugirango ikore hemolitike ya lecithine, ikora kuri selile zitukura zitanga hemolysis itaziguye.Nubwo PLA2 ari myinshi mumiryango ine yuburozi bwinzoka, ibirimo imisemburo muburozi bwinzoka buratandukanye gato.Inzoka y'inzoka (C.
Ubumara bwinzoka bwerekanaga gusa intege nke za PLA2.Imbonerahamwe 3-11-11 yerekana kugereranya ibikorwa bya PLA2 byuburozi 10 bwinzoka zifite ubumara mubushinwa.
Imbonerahamwe 3-11-11 Kugereranya ibikorwa bya fosifolipase VIII yuburozi 10 bwinzoka mubushinwa
Ubumara bwinzoka
Kurekura ibinure
Acide Alifatique,
Cjumol / mg)
Igikorwa cya Hemolytike CHU50 / ^ g * ml)
Ubumara bwinzoka
Kurekura aside irike
(^ raol / mg)
Igikorwa cya Hemolytike “(HU50 / ftg * 1111)
Najanaja atra
9. 62
cumi n'umwe
Micracephal ophis
ingingo eshanu zeru
kalyspallas
8. 68
ibihumbi bibiri na magana inani
gracilis
V, acutus
7. 56
* * #
Ophiophagus hannah
ingingo eshatu umunani ebyiri
ijana na mirongo ine
Bnugarus fasctatus
7,56
magana abiri na mirongo inani
B. multicinctus
ingingo imwe icyenda itandatu
magana abiri na mirongo inani
Viper a russelli
amanota arindwi zeru
T, mucrosquamatus
ingingo imwe umunani gatanu
Siamensis
T. stejnegeri
0. 97
（2） Gutandukana no kwezwa
Ibiri muri PLA2 muburozi bwinzoka ni binini, kandi birahamye gushyushya, aside, alkali na denaturant, kuburyo byoroshye kweza no gutandukanya PLA2.Uburyo busanzwe ni ukubanza gukora filtration ya gel ku bumara butavanze, hanyuma ugakora ion yo guhana chromatografiya, kandi intambwe ikurikira irashobora gusubirwamo.Twabibutsa ko gukonjesha-gukama kwa PLA2 nyuma ya ion-guhanahana chromatografiya bitagomba guteranya, kuko inzira yo gukonjesha akenshi yongerera imbaraga ionic muri sisitemu, kikaba ari ikintu cyingenzi giteranya PLA2.Usibye uburyo rusange bwavuzwe haruguru, uburyo bukurikira nabwo bwakoreshejwe: ① Iriba nibindi.An Substrate analogue ya PLA2 yakoreshejwe nka ligand ya affinity chromatography.Iyi ligand irashobora guhuza na PLA2 muburozi bwinzoka hamwe na Ca2 +.EDTA ikoreshwa cyane cyane.Ca2 + imaze gukurwaho, isano iri hagati ya PLA2 na ligand iragabanuka, kandi irashobora gutandukana na ligand.Abandi bakoresha 30% igisubizo kama cyangwa 6mol / L urea nkibisanzwe.Chromatografi ya Hydrophobique yakorewe hamwe na PheiiylSephar0SeCL-4B kugirango ikureho PLA2 muri cardiotoxine.Ant Antibody Anti PLA2 yakoreshejwe nka ligand kugirango ikore chromatografi ya affinity kuri PLA2.
Kugeza ubu, umubare munini w’ubumara bwinzoka PLAZ bumaze kwezwa.Tu n'abandi..Hano, twibanze ku gutandukanya no kwezwa kwa PLA nintiti zUbushinwa.
Chen Yuancong n'abandi..Ukurikije uburozi bwayo, PLA2 itabogamye ni uburozi bwinshi, bwamenyekanye nka presinaptic neurotoxin Agkistrodotoxin.Alkaline PLA2 ntabwo ifite uburozi buke, kandi aside PLA2 idafite uburozi.Wu Xiangfu n'abandi..Ibisubizo byerekanaga ko bafite uburemere buke bwa molekuline hamwe nubushyuhe bwumuriro, ariko bifite itandukaniro rikomeye mubindi bice.Mu rwego rwibikorwa bya enzyme, ibikorwa bya enzyme ya aside byari hejuru yibikorwa bya alkaline enzyme;Ingaruka ya hemolytike ya enzyme ya alkaline kuri selile yamaraso itukura niyo yari ikomeye cyane, ikurikirwa na enzyme idafite aho ibogamiye, hamwe na enzyme ya aside irike.Kubwibyo, haravugwa ko ingaruka ya hemolytic ya PLAZ ifitanye isano no kwishyuza molekile ya PLA2.Zhang Jingkang n'abandi.(1981) bakoze kristu ya Agkistrodotoxin.Tu Guangliang n'abandi.. -terminal yagenwe.Li Yuesheng n'abandi.(1985) yitaruye kandi yeza indi PLA2 mu burozi bwa Viper rotundus muri Fujian.Subunit ya PLA2 * ni 13 800, ingingo ya isoelectric ni 10.4, kandi ibikorwa byihariye ni 35 / xnioI / miri mg。 Hamwe na lecithine nka substrate, pH nziza ya enzyme ni 8.0 naho ubushyuhe bwiza ni 65 ° C. LD5 yatewe inshinge mu mbeba.Ni 0.5 ± 0.12mg / kg.Iyi misemburo ifite anticoagulant igaragara na hemolytike.Molekile yubumara ya PLA2 igizwe nibisigara 123 byubwoko 18 bwa aside amine.Molekile ikungahaye kuri sisitemu (14), aside aside (14) na glycine (12), ariko irimo methionine imwe gusa, kandi N-terminal yayo ni ibisigarira bya serine.Ugereranije na PLA2 yitandukanije na Tuguang, uburemere bwa molekile n'umubare w'ibisigisigi bya aside amine ya isoenzymes zombi birasa cyane, kandi ibigize aside amine nabyo birasa cyane, ariko umubare wa acide aspartic hamwe n'ibisigazwa bya protine biratandukanye.Guangxi king cobra ubumara bwinzoka burimo PLA2 ikize.Shu Yuyan n'abandi..Duhereye kuri microscope ya electron yerekana ingaruka za PLA2 yibanze kuri selile yamaraso itukura, urashobora kubona ko igira ingaruka zigaragara kumitsi itukura yumuntu utukura, ariko nta ngaruka zigaragara zifite mumaraso atukura y'ihene.Iyi PLA2 ifite ingaruka zo kudindiza umuvuduko wa electrophoreque ya selile yamaraso itukura mubantu, ihene, inkwavu ningurube.Chen n'abandi.Iyi misemburo irashobora kubuza gukusanya platelet iterwa na ADP, kolagen na sodium arachidonic.Iyo kwibanda kwa PLA2 ari 10 / xg / ml ~ lOOjug / ml, guteranya platine birabujijwe rwose.Niba platine yogejwe yakoreshejwe nkibikoresho, PLA2 ntishobora kubuza kwegeranya kuri 20Mg / ml.Aspirine ni inzitizi ya cyclooxygenase, ishobora kubuza ingaruka za PLA2 kuri platine.PLA2 irashobora kubuza gukusanya platine ukoresheje hydrolyzing acide arachidonic kugirango ihuze tromboxane A2.Guhindura igisubizo cya PLA2 byakozwe na Agkistrodon halys Pallas ubumara bwa Pallas mu Ntara ya Zhejiang byakozweho ubushakashatsi hakoreshejwe dichroism izenguruka, fluorescence hamwe no kwinjiza UV.Ibisubizo byubushakashatsi byagaragaje ko ihinduka nyamukuru ryuru ruhererekane rwiyi misemburo rwasaga nubwoko bumwe bwa enzyme yo mu yandi moko na genera, ihinduka rya skeleton ryarwanije ubushyuhe bwiza, kandi ihinduka ryimiterere mubidukikije bya aside ntirishobora guhinduka.Gukomatanya kwa activateur Ca2 + na enzyme ntabwo bigira ingaruka kubidukikije bya tryptophan, mugihe inhibitor Zn2 + ikora ibinyuranye.Uburyo pH agaciro k'igisubizo bigira ingaruka kubikorwa bya enzyme bitandukanye na reagent yavuzwe haruguru.
Mubikorwa byo kweza PLA2 uburozi bwinzoka, ikintu kigaragara nuko ubumara bwinzoka burimo impinga ebyiri cyangwa nyinshi zo gukuraho PLA2.Iki kintu gishobora gusobanurwa gutya: ① bitewe no kubaho kwa isozymes;Ubwoko bumwe bwa PLA2 bwahinduwe muburyo butandukanye bwa PLA2 buvanze nuburemere butandukanye bwa molekile, inyinshi murizo ziri hagati ya 9 000 ~ 40 000;③ Guhuza PLA2 nibindi bigize ubumara bwinzoka bigora PLA2;④ Kuberako inkwano ya amide muri PLA2 iba hydrolyzed, amafaranga arahinduka.① Kandi ② birasanzwe, usibye bike, nka PLA2 muburozi bwinzoka CrWa / w
Hariho ibintu bibiri: ① na ②.Indwara ya gatatu yabonetse muri PLA2 muburozi bwinzoka zikurikira: Oxyranus scutellatus, microlepidota ya Parademansia, Bothrops a ^> er, inzoka ya Palesitine, inzoka zumusenyi, ninzoka iteye ubwoba km。
Ibisubizo byurubanza ④ bituma umuvuduko wimuka wa PLA2 uhinduka mugihe cya electrophore, ariko aside amine ntabwo ihinduka.Peptide irashobora kumenwa na hydrolysis, ariko mubisanzwe iracyahujwe hamwe na disulfide.Ubumara bwinzoka yinzoka yiburasirazuba burimo uburyo bubiri bwa PLA2, bwitwa ubwoko a nubwoko bwa p PLA2.Itandukaniro riri hagati yubwoko bubiri bwa PLA2 ni aside amine imwe gusa, ni ukuvuga glutamine muri molekile imwe ya PLA2 isimburwa na acide glutamic mubindi molekile ya PLA2.Nubwo impamvu nyayo y'iri tandukaniro idasobanutse, muri rusange abantu bemeza ko ifitanye isano no gusenya PLA2.Niba PLA2 muburozi bwa viper yo muri Palesitine ikomeje gushyuha hamwe nuburozi bubi, amatsinda yanyuma muri molekile ya enzyme yayo azaba menshi kurusha mbere.Kuva kuri C PLA2 yitandukanije nuburozi bwinzoka ifite N-terminal ebyiri zitandukanye, kandi uburemere bwa molekile ni 30000. Iki kintu gishobora guterwa na dimim asimmetrike ya PLA2, isa na dimimmetrike yakozwe na PLA2 muburozi bwinzoka ya diyama yiburasirazuba. n'inzoka ya diamback yiburengerazuba.Cobra yo muri Aziya igizwe nubwoko butandukanye, bumwe murubwo ntibusobanutse neza mubyiciro.Kurugero, icyahoze cyitwa Cobra Outer Caspian subcecies ubu kiramenyekana
Bikwiye kwitirirwa Cobra yo mu nyanja ya Kaspiya.Nkuko hariho amoko menshi kandi akavangwa hamwe, ubumara bwinzoka buratandukanye cyane kubera amasoko atandukanye, kandi nibiri muri isozymes ya PLA2 nabyo ni byinshi.Kurugero, ubumara bwa cobra
Nibura ubwoko 9 bwa isozymes ya PLA2 yubwoko bwa r ^ ll bwabonetse, naho ubwoko 7 bwa isozymes ya PLA2 bwabonetse muburozi bwubwoko bwa cobra Caspian.Durkin n'abandi..Muri rusange, ibikorwa bya PLA2 byuburozi bwa cobra ni byinshi, hamwe na isozymes nyinshi.Igikorwa cya PLA2 na isozymes yuburozi bwa viper ni buke.Ibikorwa bya PLA2 byuburozi bwa mamba nuburozi bwinzoka ni bike cyane cyangwa nta gikorwa cya PLA2.Ibikorwa bya PLA2 byuburozi bwinzoka zo mu nyanja nabyo ni bike.
Mu myaka yashize, ntabwo byavuzwe ko PLA2 mu bumara bwinzoka ibaho muburyo bwa dimer ikora, nk'inzoka ya rhombophora y'iburasirazuba (C. ubumara bwinzoka burimo ubwoko bwubwoko bwa P PLA2, byombi bigizwe na subunits ebyiri zisa. , na dimerase gusa ifite
Igikorwa.Shen n'abandi.yasabye kandi ko dimer ya PLA2 yuburozi bwinzoka aribwo buryo bukora bwa enzyme.Ubushakashatsi bwimiterere yimiterere burerekana kandi ko PLA2 yinzoka ya diyama yiburengerazuba ibaho muburyo bwa dimer.Piscivorous compound
Hariho PLA ebyiri zitandukanye ^ Ei na E2 z'ubumara bwinzoka, aho 仏 ibaho muburyo bwa dimer, dimer irakora, kandi monomer yatandukanijwe idakora.Lu Yinghua n'abandi.(1980) yarushijeho kwiga imiterere yumubiri nubumara hamwe na reaction ya E. Jayanthi nabandi.(1989) yatandukanije PLA2 shingiro (VRVPL-V) nuburozi bwinzoka.Uburemere bwa molekuline ya monomer PLA2 ni 10000, ifite ingaruka zica, anticoagulant na edema.Enzyme irashobora gukora polymerime ifite uburemere butandukanye bwa molekuline bitewe na PH 4.8, kandi urugero rwa polymerisation hamwe nuburemere bwa molekile ya polymers byiyongera hamwe nubushyuhe bwiyongera.Uburemere bwa molekuline ya polymer yakozwe kuri 96 ° C ni 53 100, naho ibikorwa bya PLA2 byiyi polymer byiyongera kubiri
Igihe cyo kohereza: Ugushyingo-18-2022