Venom nke agwọ nwere enzymes na-eme ka njikọ nke carboxyl ester hydrolyze.Ihe mejupụtara maka hydrolysis bụ phospholipids, acetylcholine na aromatic acetate.Enzymes ndị a gụnyere ụdị atọ: phospholipase, acetylcholinesterase na aromatic esterase.Arginine esterase na venom agwọ nwekwara ike hydrolyze synthetic arginine ma ọ bụ lysine, ma ọ na-abụkarị hydrolyzes protein peptide bond na okike, ya mere ọ bụ nke protease.Enzymes ndị a tụlere ebe a na-arụ ọrụ naanị na mkpụrụ ester na enweghị ike ịme ihe na njikọ peptide ọ bụla.N'ime enzymes ndị a, ọrụ ndu nke acetylcholinesterase na phospholipase dị mkpa karịa ma mụọ ya nke ọma.Ụfọdụ venoms agwọ nwere ọrụ esterase aromatic siri ike, nke nwere ike hydrolyze p-nitrophenyl ethyl ester, a - ma ọ bụ P-naphthalene acetate na indole ethyl ester.A ka amabeghị ma ọrụ a sitere na enzyme nọọrọ onwe ya ma ọ bụ mmetụta akuku amaara nke carboxylesterase, ma ọbụghị uru ọ bara.Mgbe venom nke Agkistrodon halys Japonicus meghachiri omume na p-nitrophenyl ethyl ester na indole ethyl ester, a hụghị hydrolysates nke p-nitrophenol na indole phenol;N'ụzọ megidere nke ahụ, ọ bụrụ na esters ndị a meghachi omume na ụdị cobra Zhoushan agwọ na venom Bungarus multicinctus agwọ, a ga-agbaze ngwa ngwa.A maara na venoms cobra ndị a nwere ọrụ cholinesterase siri ike, nke nwere ike ibute hydrolysis nke ihe ndị dị n'elu.N'ezie, Mclean et al.(1971) kọrọ na ọtụtụ nsị agwọ nke ezinụlọ Cobra nwere ike mee ka indole ethyl ester, naphthalene ethyl ester na butyl naphthalene ester mee mmiri.Anụ agwọ ndị a na-esi: cobra, olu olu ojii, agụba egbugbere ọnụ ojii, ugo ọla edo, ụgbụgba Ijipt, ugo eze, ụgbụgba ọla edo, mamba ojii na ọcha egbugbere ọnụ mamba (D. aw ka maara rhombola rattlesnake ọwụwa anyanwụ.
Venom agwọ nwere ike hydrolyze methyl indole ethyl ester, nke bụ mkpụrụ maka ịchọpụta ọrụ cholinesterase na serum, mana venom agwọ a anaghị egosi ọrụ cholinesterase.Nke a na-egosi na e nwere ihe esterase a na-amaghị ama na venom cobra, nke dị iche na cholinesterase.Iji ghọta ọdịdị nke enzyme a, a chọrọ ọrụ nkewa ọzọ.
1, Phospholipase A2
(I) Nchịkọta
Phospholipase bụ enzyme nke nwere ike imebi glyceryl phosphate.Enwere ụdị phospholipase 5 na okike, ya bụ phospholipase A2 na phospholipase.
A. , phospholipase B, phospholipase C na phospholipase D. Nri agwọ na-abụkarị phospholipase A2 (PLA2), nsị agwọ ole na ole nwere phospholipase B, na phospholipases ndị ọzọ na-ahụkarị na anụ ahụ na nje bacteria.Fig. 3-11-4 na-egosi ebe a na-eme ihe nke phospholipases ndị a na mkpụrụ osisi hydrolysis.
N'ime phospholipases, a mụọla PLA2 karịa.O nwere ike ịbụ enzyme kacha mụọ na nsị agwọ.Mkpụrụ ya bụ njikọ ester na ọnọdụ nke abụọ nke Sn-3-glycerophosphate.A na-ahụ enzyme a n'ọtụtụ ebe na nsị agwọ, venom bee, venom scorpion na anụ ahụ anụmanụ, na PLA2 dị ukwuu n'ime anụ agwọ ezinụlọ anọ.N'ihi na enzyme a na-agbaji mkpụrụ ndụ ọbara uhie ma na-ebute hemolysis, a na-akpọkwa ya "hemolysin".Ụfọdụ ndị mmadụ na-akpọkwa PLA2 hemolytic lecithinase.
Ludeeke buru ụzọ chọpụta na venom agwọ nwere ike imepụta ogige hemolytic site n'ime ihe lecithin site na enzymes.Mgbe e mesịrị, Delezenne et al.gosiputara na mgbe venom cobra na-eme na serum inyinya ma ọ bụ nkochi ime akwa, ọ na-etolite ihe hemolytic.A maara ugbu a na PLA2 nwere ike ime kpọmkwem na phospholipids nke akpụkpọ ahụ erythrocyte, na-ebibi usoro nke akpụkpọ ahụ erythrocyte ma na-akpata hemolysis ozugbo;Ọ nwekwara ike ime na serum ma ọ bụ tinye lecithin iji mepụta lecithin hemolytic, nke na-arụ ọrụ na mkpụrụ ndụ ọbara uhie iji mepụta hemolysis na-apụtaghị ìhè.Ọ bụ ezie na PLA2 dị ukwuu n'ezinụlọ anọ nke nsị agwọ, ọdịnaya nke enzymes dị na nsị agwọ dị iche iche dịtụ iche.Rattlesnake (C
Nri agwọ gosipụtara naanị ọrụ PLA2 adịghị ike.Tebụl 3-11-11 na-egosi atụnyere ọrụ PLA2 nke isi 10 nke agwọ ọjọọ na China.
Tebụl 3-11-11 atụnyere ihe omume phospholipase nke Asatọ nke nsị agwọ iri na China.
Egwu agwọ
Ntọhapụ abụba
Aliphatic acid,
Cjumol/mg)
Ọrụ hemolytic CHU50/^ g * ml)
Egwu agwọ
Hapụ acid fatty
(^raol/mg)
Ọrụ hemolytic “(HU50/ftg * 1111)
Najanaja atra
9.62
iri na otu
Microcephalo ophis
isi ise otu efu
kalyspallas
8.68
puku abụọ na narị asatọ
gracilis
V, akutus
7.56
* * #
Ophiophagus hannah
isi atọ asatọ abụọ
otu narị na iri anọ
Bnugarus fatịlaịza
7,56
narị abụọ na iri asatọ
B. multicinctus
otu isi itoolu isii
narị abụọ na iri asatọ
Viper na russelli
akara asaa efu atọ
T, mucrosquamatus
otu isi asatọ ise
Siamensis
T. stejnegeri
0.97
(2) Nkewa na ịdị ọcha
Ọdịnaya nke PLA2 na venom agwọ buru ibu, ọ na-akwụsikwa ike ikpo ọkụ, acid, alkali na denaturant, nke mere na ọ dị mfe ime ka ọ dị ọcha na ikewapụ PLA2.Usoro a na-emekarị bụ ibu ụzọ mee gel filtration na venom crude, wee mee ion mgbanwe chromatography, na nzọụkwụ ọzọ nwere ike ime ugboro ugboro.Ekwesiri ighota na ikpochapu-ihicha nke PLA2 mgbe ion-exchange chromatography ekwesịghị ime ka nchịkọta, n'ihi na usoro ikpochapu na-eme ka ike ion na-abawanye na usoro ahụ, nke bụ ihe dị mkpa na-eme ka nchịkọta nke PLA2.Na mgbakwunye na usoro izugbe ndị dị n'elu, a nabatakwara ụzọ ndị a: ① Wells et al.② A na-eji ihe analog nke PLA2 mee ihe dị ka ligand maka chromatography mmekọrịta.Njikọ a nwere ike jikọta PLA2 na nsị agwọ na Ca2+.A na-ejikarị EDTA eme ihe dị ka eluent.Mgbe ewepụrụ Ca2 +, mmekọrịta dị n'etiti PLA2 na ligand na-ebelata, enwere ike kewapụ ya na ligand.Ndị ọzọ na-eji 30% organic solution ma ọ bụ 6mol/L urea dị ka eluent.③ Ejiri PheiiylSephar0SeCL-4B mee chromatography hydrophobic iji wepu akara PLA2 na cardiotoxin.④ A na-eji ọgwụ mgbochi PLA2 eme ihe dị ka ligand iji mee chromatography mmekọrịta na PLA2.
Ka ọ dị ugbu a, a sachapụla ọnụ ọgụgụ buru ibu nke venom agwọ.Tu et al.(1977) depụtara PLA2 dị ọcha site na venom agwọ tupu 1975. N'afọ ndị na-adịbeghị anya, a na-akọ ọnụ ọgụgụ buru ibu nke akụkọ gbasara nkewa na ịdị ọcha nke PLA2 kwa afọ.N'ebe a, anyị na-elekwasị anya na nkewa na ịdị ọcha nke PLA site n'aka ndị ọkà mmụta China.
Chen Yuancong et al.(1981) kewapụrụ ụdị PLA2 atọ na venom nke Agkistrodon halys Pallas na Zhejiang, nke nwere ike kewaa n'ime acidic, nnọpụiche na alkaline PLA2 dị ka isi ihe isoelectric ha si dị.Dị ka nsi ya si dị, PLA2 na-anọpụ iche bụ ihe na-egbu egbu karị, nke achọpụtara dị ka presynaptic neurotoxin Agkistrodotoxin.Alkaline PLA2 adịghị egbu egbu, yana PLA2 acidic enweghị ihe fọrọ nke nta ka ọ bụrụ nsí.Wu Xiangfu et al.(1984) tụnyere njirimara nke PLA2 atọ, gụnyere ịdị arọ molecular, amino acid mejupụtara, N-terminal, isoelectric point, nkwụsi ike nke okpomọkụ, ọrụ enzyme, nsị na ọrụ hemolytic.Nsonaazụ gosiri na ha nwere otu ịdị arọ molekụla na nkwụsi ike ọkụ, mana ha nwere nnukwu ọdịiche na akụkụ ndị ọzọ.N'akụkụ nke ọrụ enzyme, ọrụ enzyme acid dị elu karịa ọrụ enzyme alkaline;Mmetụta hemolytic nke enzyme alkaline na mkpụrụ ndụ ọbara uhie oke bụ nke kachasị ike, na-esote enzyme na-anọpụ iche, na enzyme acid na-esighi ike hemolyzed.Ya mere, a na-eche na mmetụta hemolytic nke PLAZ metụtara ụgwọ nke molekul PLA2.Zhang Jingkang et al.(1981) emeela Agkistrodotoxin kristal.Tu Guangliang et al.(1983) kọrọ na PLA na-egbu egbu nke nwere ebe isoelectric nke 7. 6 dịpụrụ adịpụ ma sachaa ya site na venom nke Vipera rotundus sitere na Fujian, yana ihe anụ ahụ na kemịkal ya, ihe mejupụtara amino acid na usoro nke 22 amino acid resides na N. -Achọpụtara ọnụ.Li Yuesheng et al.(1985) kewapụrụ ma sachaa PLA2 ọzọ site na venom nke Viper rotundus na Fujian.Akụkụ nke PLA2 * bụ 13 800, isi ihe isoelectric bụ 10.4, yana ọrụ a kapịrị ọnụ bụ 35/xnioI/miri mg. Agbanye LD5 n'ime ụmụ oke.Ọ bụ 0.5 ± 0.12mg / kg.Enzyme a nwere mmetụta anticoagulant doro anya na mmetụta hemolytic.Ngwakọta PLA2 na-egbu egbu nwere ihe fọdụrụ 123 nke ụdị amino acid iri na asatọ.Ngwakọta ahụ bara ụba na cysteine (14), aspartic acid (14) na glycine (12), mana ọ nwere naanị otu methionine, yana N-terminal ya bụ ihe fọdụrụ na serine.E jiri ya tụnyere PLA2 dịpụrụ adịpụ site na Tuguang, ịdị arọ molekụla na ọnụ ọgụgụ amino acid residues nke isoenzymes abụọ yiri nke ahụ, na amino acid mejupụtara nwekwara yikọrọ, ma ọnụ ọgụgụ nke aspartic acid na proline residues dịtụ iche.Venom nke eze cobra nke Guangxi nwere PLA2 bara ụba.Shu Yuyan et al.(1989) dịpụrụ adịpụ PLA2 site na venom, nke nwere otu ọrụ 3.6 ugboro dị elu karịa venom mbụ, ịdị arọ molecular nke 13000, ihe mejupụtara 122 amino acid residues, ebe isoelectric nke 8.9, na ezigbo nkwụsi ike nke thermal.Site na nyocha microscope eletrọn nke mmetụta PLA2 bụ isi na mkpụrụ ndụ ọbara uhie, enwere ike ịhụ na ọ nwere mmetụta doro anya na akpụkpọ anụ ọbara ọbara mmadụ, mana enweghị mmetụta doro anya na mkpụrụ ndụ ọbara uhie ewu.Nke a PLA2 nwere mmetụta nkwụghachi azụ doro anya na ọsọ electrophoretic nke mkpụrụ ndụ ọbara uhie na mmadụ, ewu, oke bekee na ezi pigs.Chen et al.Enzyme a nwere ike igbochi nchịkọta platelet nke ADP, collagen na sodium arachidonic acid kpatara.Mgbe ntinye PLA2 bụ 10/xg/ml~lOOjug/ml, a na-egbochi nchịkọta platelet kpamkpam.Ọ bụrụ na ejiri platelet a sachara mee ihe dị ka ihe, PLA2 enweghị ike igbochi mkpokọta na mkpokọta 20Mg / ml.Aspirin bụ onye na-egbochi cyclooxygenase, nke nwere ike igbochi mmetụta PLA2 na platelet.PLA2 nwere ike igbochi nchịkọta platelet site na hydrolyzing arachidonic acid iji mepụta thromboxane A2.Ngwọta ngwọta nke PLA2 nke Agkistrodon halys Pallas venom mepụtara na mpaghara Zhejiang ka a mụrụ site na dichroism okirikiri, fluorescence na absorption nke UV.Nsonaazụ nnwale gosiri na isi ihe ngbanwe agbụ nke enzyme a yiri nke otu ụdị enzyme sitere na ụdị ndị ọzọ na mkpụrụ ndụ ihe nketa, ọkpụkpụ ọkpụkpụ nwere ezigbo nguzogide okpomọkụ, mgbanwe nhazi na gburugburu acid nwere ike gbanwee.Nchikota nke activator Ca2 + na enzyme anaghị emetụta gburugburu nke ihe fọdụrụ tryptophan, ebe onye na-egbochi Zn2 + na-eme ihe dị iche.Ụzọ nke uru pH nke ngwọta na-emetụta ọrụ enzyme dị iche na reagents dị n'elu.
N'ime usoro nke PLA2 dị ọcha nke nsị agwọ, ihe pụtara ìhè bụ na nsị agwọ nwere elu elu abụọ ma ọ bụ karịa PLA2.Enwere ike ịkọwa ihe omume a dị ka ndị a: ① n'ihi ịdị adị nke isozymes;② Otu ụdị PLA2 bụ polymerized n'ime ụdị PLA2 dị iche iche nke nwere ihe dị iche iche molekụla, ọtụtụ n'ime ha dị na 9 000 ~ 40 000;③ Ngwakọta PLA2 na ihe ndị ọzọ na-eme nsị agwọ na-eme ka PLA2 sie ike;④ N'ihi na njikọ amide dị na PLA2 bụ hydrolyzed, ụgwọ ahụ na-agbanwe.① Na ② bụ ihe a na-ahụkarị, ma ewezuga ole na ole, dị ka PLA2 na venom agwọ CrWa/w
Enwere ọnọdụ abụọ: ① na ②.Achọtala ọnọdụ nke atọ na PLA2 na nsị nke agwọ ndị a: Oxyranus scutellatus, Parademansia microlepidota, Bothrops a ^>er, viper Palestine, viper ájá, na egwu rattlesnake km.
Nsonaazụ nke ikpe ④ na-eme ka mgbanwe ọsọ nke PLA2 gbanwee n'oge electrophoresis, mana amino acid anaghị agbanwe agbanwe.Enwere ike imebi peptide site na hydrolysis, mana n'ozuzu ha ka na-ejikọta ya site na njikọ disulfide.Venom nke olulu mmiri ọwụwa anyanwụ nwere ụdị abụọ nke PLA2, nke a na-akpọ ụdị a na ụdị p PLA2 n'otu n'otu.Ọdịiche dị n'etiti ụdị PLA2 abụọ a bụ naanị otu amino acid, ya bụ, glutamine n'ime otu molecule PLA2 na-eji glutamic acid dochie ya na molecule PLA2 ọzọ.Ọ bụ ezie na kpọmkwem ihe kpatara ọdịiche a abụghị nke doro anya, a na-ekwenyekarị na ọ na-ejikọta ya na deamination nke PLA2.Ọ bụrụ na PLA2 na venom nke Palestine na-ekpo ọkụ na venom crude, otu njedebe na mkpụrụ ndụ enzyme ya ga-adị karịa ka ọ dị na mbụ.Site na C PLA2 dịpụrụ adịpụ site na venom agwọ nwere N-terminal abụọ dị iche iche, na ịdị arọ ya bụ 30000. Ihe a nwere ike ime site na asymmetric dimer nke PLA2, nke yiri symmetric dimer nke PLA2 mere na venom nke ọwụwa anyanwụ diamondback rattlesnake. na ọdịda anyanwụ diamondback rattlesnake.Ihe mejupụtara ube Asia nwere ọtụtụ ụdị, ụfọdụ n'ime ha amachaghị nke ọma na nhazi.Dịka ọmụmaatụ, a matala ihe a na-akpọbu ngalaba Cobra Outer Caspian ugbu a
Ekwesịrị ịsị na ọ bụ Oke Osimiri Caspian dị n'èzí.Ebe ọ bụ na e nwere ọtụtụ subspecies na ha na-agwakọta ọnụ, mejupụtara venom agwọ dịgasị iche iche nke ukwuu n'ihi na dị iche iche isi mmalite, na ọdịnaya nke PLA2 isozymes dịkwa elu.Dịka ọmụmaatụ, nsị uka
Opekempe, a chọtara ụdị 9 dị iche iche nke PLA2 isozymes nke ụdị r ll, na ụdị 7 dị iche iche nke PLA2 isozymes ka ahụrụ n'ime venom nke subspecies cobra Caspian.Durkin et al.(1981) mụọ ọdịnaya PLA2 na ọnụ ọgụgụ nke isozymes dị na nsị agwọ dị iche iche, gụnyere 18 venoms cobra, 3 mamba venoms, 5 viper venoms, 16 rattlesnake venoms na 3 venoms agwọ oké osimiri.N'ozuzu, ọrụ PLA2 nke venom cobra dị elu, nwere ọtụtụ isozymes.Ọrụ PLA2 na isozymes nke viper venom bụ ọkara.Omume PLA2 nke mamba venom na rattlesnake venom dị obere ma ọ bụ enweghị ọrụ PLA2.Ọrụ PLA2 nke nsị agwọ oké osimiri dịkwa ala.
N'afọ ndị na-adịbeghị anya, a kọwabeghị na PLA2 na nsị agwọ dị n'ụdị dimer na-arụ ọrụ, dị ka rhombophora rattlesnake nke ọwụwa anyanwụ (C. venom agwọ nwere ụdị a na ụdị PLA2, ha abụọ mejupụtara subunits abụọ yiri ya. , na naanị dimerase nwere
Ihe omume.Shen et al.tụkwara aro na naanị dimer nke PLA2 nke venom agwọ bụ ụdị arụ ọrụ nke enzyme ahụ.Ọmụmụ ihe gbasara oghere na-egosikwa na PLA2 nke ọdịda anyanwụ diamondback rattlesnake dị n'ụdị dimer.Ngwakọta piscivorous
Enwere PLA ^ Ei na E2 abụọ dị iche iche nke nsị agwọ, nke 仏 dị n'ụdị dimer, dimer na-arụ ọrụ, na monomer ya kewapụrụ adịghị arụ ọrụ.Lu Yinghua et al.(1980) gara n'ihu mụọ ihe gbasara anụ ahụ na kemịkalụ na mmeghachi omume kinetics nke E. Jayanthi et al.(1989) kewapụrụ isi PLA2 (VRVPL-V) site na venom viper.Ibu molekụla nke monomer PLA2 bụ 10000, nke nwere mmetụta na-egbu egbu, anticoagulant na edema.Enzyme nwere ike polymerize polymers dị iche iche molekụla arọ n'okpuru ọnọdụ nke PH 4.8, na ogo nke polymerization na molekụla arọ nke polymers na-abawanye na elu okpomọkụ.Ihe dị arọ nke polymer emepụtara na 96 Celsius C bụ 53 100, yana ọrụ PLA2 nke polymer a na-abawanye site na abụọ.
Oge nzipu: Nov-18-2022