Loaʻa i ka ʻawaʻawa o ka nahesa nā enzyme e hoʻoheheʻe i nā paʻa carboxyl ester.ʻO nā substrate no ka hydrolysis he phospholipids, acetylcholine a me ka acetate aromatic.Aia kēia mau enzymes i ʻekolu ʻano: phospholipase, acetylcholinesterase a me aromatic esterase.Hiki i ka Arginine esterase i loko o ka snake venom ke hydrolyze i ka arginine synthetic a i ʻole lysine, akā ʻo ia ka hydrolyzes i nā mea paʻa peptide protein i ke ʻano, no laila no ka protease.ʻO nā enzymes i kūkākūkā ʻia ma ʻaneʻi e hana wale ma luna o nā substrates ester a ʻaʻole hiki ke hana i kekahi paʻa peptide.Ma waena o kēia mau enzymes, ʻoi aku ka nui o nā hana biological o acetylcholinesterase a me phospholipase a ua aʻo piha ʻia.Loaʻa i kekahi mau mea ʻawaʻawa ʻawaʻawa ikaika ka hana esterase aromatic, hiki iā ia ke hydrolyze p-nitrophenyl ethyl ester, a - a i ʻole P-naphthalene acetate a me indole ethyl ester.ʻAʻole ʻike ʻia inā hana ʻia kēia haʻawina e kahi enzyme kūʻokoʻa a i ʻole ka hopena ʻaoʻao o ka carboxylesterase, e waiho wale i kona ʻano koʻikoʻi.I ka wā i hana ʻia ai ka mea make o Agkistrodon halys Japonicus me ka p-nitrophenyl ethyl ester a me ka indole ethyl ester, ʻaʻole i loaʻa nā hydrolysates o p-nitrophenol a me indole phenol;ʻO ka mea ʻē aʻe, inā e hana kēia mau esters me ka cobra Zhoushan subspecies snake venom a me Bungarus multicinctus snake venom, e hoʻokahe koke ʻia lākou.Ua ʻike ʻia he hana cholinesterase ikaika kēia mau cobra, ʻo ia paha ke kuleana no ka hydrolysis o nā substrates ma luna.ʻOiaʻiʻo, ʻo Mclean et al.(1971) hōʻike i ka nui o nā mea make nahesa no ka ʻohana Cobra hiki ke hoʻoheheʻe i ka indole ethyl ester, naphthalene ethyl ester a me butyl naphthalene ester.No loko mai o kēia mau mea ʻawaʻawa nahesa: cobra, ʻāʻī ʻeleʻele, cobra lehelehe ʻeleʻele, cobra gula, cobra ʻAigupita, kiʻi cobra, mamba gula, mamba ʻeleʻele a me ka mamba lehelehe keʻokeʻo (D. aw still knows eastern rhombola rattlesnake
Hiki i ka snake venom ke hoʻokahe i ka methyl indole ethyl ester, ʻo ia ka substrate no ka hoʻoholo ʻana i ka hana cholinesterase i loko o ka serum, akā ʻaʻole hōʻike kēia snake venom i ka hana cholinesterase.Hōʻike kēia aia kahi esterase ʻike ʻole i loko o ka cobra venom, he ʻokoʻa ia me ka cholinesterase.No ka hoʻomaopopo ʻana i ke ʻano o kēia enzyme, pono ka hana hoʻokaʻawale hou aku.
1、 Phospholipase A2
(I) Nānā
ʻO Phospholipase kahi enzyme e hiki ke hoʻokahe i ka glyceryl phosphate.He 5 ʻano phospholipase ma ke ʻano, ʻo ia ka phospholipase A2 a me ka phospholipase.
A. , phospholipase B, phospholipase C a me phospholipase D. ʻO ka mea make o ka nahesa ka nui o ka phospholipase A2 (PLA2), kekahi mau mea make nahesa he phospholipase B, a ʻo nā phospholipase ʻē aʻe i loaʻa nui i loko o nā ʻiʻo holoholona a me nā bacteria.Hōʻike ka Fig. 3-11-4 i ke kahua hana o kēia mau phospholipases ma ka substrate hydrolysis.
Ma waena o nā phospholipases, ua aʻo hou ʻia ka PLA2.ʻO ia paha ka enzyme i aʻo nui ʻia i ka make o ka nahesa.ʻO kona substrate ka paʻa ester ma ke kūlana ʻelua o Sn-3-glycerophosphate.Loaʻa nui ʻia kēia enzyme i loko o ka snake venom, bee venom, scorpion scorpion a me nā ʻiʻo holoholona, a nui ka PLA2 i loko o ʻehā ʻohana nahesa.Ma muli o ka haki ʻana o kēia enzyme i ke koko ʻulaʻula a me ka hemolysis, ua kapa ʻia ʻo ia ʻo "hemolysin".Kāhea kekahi poʻe iā PLA2 hemolytic lecithinase.
Ua ʻike mua ʻo Ludeeke e hiki i ka nahesa ke hana i kahi pūhui hemolytic ma ka hana ʻana i ka lecithin ma o nā enzymes.Ma hope mai, ʻo Delezenne et al.ua hōʻike ʻia i ka wā e hana ai ka make ʻawaʻawa ma luna o ka serum lio a i ʻole yolk, lilo ia i mea hemolytic.Ua ʻike ʻia i kēia manawa hiki i ka PLA2 ke hana pololei i nā phospholipids o ka membrane erythrocyte, e luku ana i ke ʻano o ka membrane erythrocyte a me ka hemolysis pololei;Hiki iā ia ke hana i ka serum a i ʻole lecithin i hoʻohui ʻia e hana i ka lecithin hemolytic, e hana ana i nā ʻulaʻula ʻulaʻula e hana i ka hemolysis indirect.ʻOiai ka nui o ka PLA2 i loko o nā ʻohana ʻehā o nā mea ʻawaʻawa nahesa, ʻokoʻa iki ka ʻike o nā enzymes i loko o nā ʻupena snake like ʻole.Nahesa (C
Ua hōʻike wale ʻia ka hana PLA2 nāwaliwali o ka nahesa.Hōʻike ka papa 3-11-11 i ka hoʻohālikelike ʻana o ka hana PLA2 o 10 mau mea make nui o nā nahesa make ma Kina.
Papa 3-11-11 Hoʻohālikelike i nā hana phospholipase VIII o 10 mau mea make nahesa ma Kina.
ʻawaʻawa nahesa
Hoʻokuʻu momona
ʻO ka waikawa aliphatic,
Cjumol/mg)
Hana hemolytic CHU50/^ g * ml)
ʻawaʻawa nahesa
Hoʻokuʻu i nā ʻakika momona
(^raol/mg)
Hana hemolytic “(HU50/ftg * 1111)
Najanaja atra
9. 62
ʻumikumākahi
ʻO Micracephal ophis
ʻelima helu hoʻokahi zero
kalyspallas
8. 68
elua tausani me ewalu haneri
gracilis
V, acutus
7. 56
* * #
Ophiophagus hannah
ʻekolu helu ʻewalu ʻelua
hookahi haneri a me kanaha
Bnugarus fasctatus
7,56
elua haneri me kanawalu
B. multicinctus
hoʻokahi helu ʻeiwa ʻeono
elua haneri me kanawalu
Viper a russelli
ʻehiku kiko ʻole ʻekolu
T, mucrosquamatus
hookahi helu ewalu elima
Siamensis
T. stejnegeri
0. 97
（2） Hoʻokaʻawale a hoʻomaʻemaʻe
ʻO ka maʻiʻo o PLA2 i ka snake venom he nui, a paʻa i ka wela, ka waikawa, ka alkali a me ka denaturant, i maʻalahi ka hoʻomaʻemaʻe a hoʻokaʻawale iā PLA2.ʻO ke ʻano maʻamau ka hana mua ʻana i ka kānana gel ma luna o ka mea ʻawaʻawa, a laila e hoʻokō i ka chromatography hoʻololi ion, a hiki ke hana hou i ka hana aʻe.Pono e hoʻomaopopo ʻia ʻaʻole pono ka hoʻomaloʻo ʻana o ka PLA2 ma hope o ka chromatography hoʻololi ion-exchange, no ka mea, ʻo ke kaʻina hoʻomaloʻo manuahi e hoʻonui pinepine ai i ka ikaika ionic i loko o ka ʻōnaehana, he kumu nui ia e hoʻohui ai i ka PLA2.Ma waho aʻe o nā ʻano hana maʻamau i luna, ua ʻae ʻia nā ʻano hana penei: ① Wells et al.② Ua hoʻohana ʻia ke analogue substrate o PLA2 ma ke ʻano he ligand no ka chromatography affinity.Hiki i kēia ligand ke hoʻopaʻa iā PLA2 i ka nahesa make me Ca2+.Hoʻohana nui ʻia ʻo EDTA ma ke ʻano he eluent.Ma hope o ka wehe ʻia ʻana o Ca2+, e emi ana ka pilina ma waena o PLA2 a me ka ligand, a hiki ke hoʻokaʻawale ʻia mai ka ligand.Hoʻohana ka poʻe ʻē aʻe i 30% ka hopena organik a i ʻole 6mol/L urea ma ke ʻano he eluent.③ Ua hana ʻia ka hydrophobic chromatography me PheiiylSephar0SeCL-4B e wehe i ka trace PLA2 i loko o ka cardiotoxin.④ Ua hoʻohana ʻia ka antibody anti PLA2 ma ke ʻano he ligand e hana i ka chromatography affinity ma PLA2.
I kēia manawa, ua hoʻomaʻemaʻe ʻia ka nui o nā snake venom PLAZ.Tu et al.(1977) papa inoa ʻo PLA2 i hoʻomaʻemaʻe ʻia mai ka nahesa ma mua o 1975. I nā makahiki i hala iho nei, ua hōʻike ʻia ka nui o nā ʻatikala e pili ana i ka hoʻokaʻawale ʻana a me ka hoʻomaʻemaʻe ʻana o PLA2 i kēlā me kēia makahiki.Maʻaneʻi, nānā mākou i ka hoʻokaʻawale ʻana a me ka hoʻomaʻemaʻe ʻana i ka PLA e nā loea Kina.
ʻO Chen Yuancong et al.(1981) hoʻokaʻawale ʻekolu mau ʻano PLA2 mai ka mea make o Agkistrodon halys Pallas ma Zhejiang, hiki ke hoʻokaʻawale ʻia i loko o ka acidic, neutral a me ka alkaline PLA2 e like me kā lākou isoelectric points.Wahi a kona ʻona, ʻoi aku ka ʻona o ka PLA2 kūʻokoʻa, i ʻike ʻia he presynaptic neurotoxin Agkistrodotoxin.ʻAʻole ʻawaʻawa ka alkaline PLA2, a ʻaneʻane ʻaʻohe mea ʻona o ka PLA2 acidic.Wu Xiangfu et al.(1984) hoʻohālikelike i nā hiʻohiʻona o nā PLA2 ʻekolu, me ke kaumaha molekala, ka hui ʻana o ka amino acid, N-terminal, isoelectric point, thermal stability, enzyme activity, toxicity and hemolytic activity.Ua hōʻike ʻia nā hopena he like ko lākou kaumaha molekala a me ke kūpaʻa wela, akā he ʻokoʻa koʻikoʻi ma nā ʻano ʻē aʻe.Ma ke ʻano o ka hana enzyme, ʻoi aku ka kiʻekiʻe o ka hana enzyme acid ma mua o ka hana enzyme alkaline;ʻO ka hopena hemolytic o ka enzyme alkaline ma ke koko ʻulaʻula ʻiole ka mea ikaika loa, a ukali ʻia e ka enzyme neutral, a ʻaʻole hiki ke hemolyzed ka acid acid.No laila, ua manaʻo ʻia e pili ana ka hopena hemolytic o PLAZ i ka hoʻopiʻi o ka mole PLA2.ʻO Zhang Jingkang et al.(1981) ua hana i nā kristal Agkistrodotoxin.Tu Guangliang et al.(1983) hōʻike i ka PLA meaʻawaʻawa me kahi isoelectric kiko o 7. 6 ua hoʻokaʻawale ʻia a hoʻomaʻemaʻe ʻia mai ka ʻawaʻawa o Vipera rotundus mai Fujian, a me kona mau kino kino a me kemika, ka hui amino acid a me ke kaʻina o 22 amino acid koena ma ka N. - ua hoʻoholo ʻia ka terminal.Li Yuesheng et al.(1985) hoʻokaʻawale a hoʻomaʻemaʻe i kahi PLA2 hou mai ka make o Viper rotundus ma Fujian.ʻO ka subunit o ka PLA2 * he 13 800, ka helu isoelectric he 10.4, a ʻo ka hana kikoʻī he 35/xnioI/miri mg. Me ka lecithin ma ke ʻano he substrate, ʻo ka pH maikaʻi loa o ka enzyme he 8.0 a ʻo ka mahana maikaʻi loa he 65 ° C. . Hoʻokomo ʻia ʻo LD5 i loko o nā ʻiole.He 0.5 ± 0.12mg/kg.Loaʻa kēia enzyme i nā hopena anticoagulant a hemolytic.He 123 koena o 18 mau ʻano amino acids ka mole PLA2 ʻona.He waiwai ka mole i ka cysteine (14), aspartic acid (14) a me glycine (12), akā hoʻokahi wale nō methionine, a ʻo kāna N-terminal he koena serine.Ke hoʻohālikelike ʻia me ka PLA2 i hoʻokaʻawale ʻia e Tuguang, ʻo ke kaumaha o ka molecular a me ka helu o nā koena amino acid o nā isoenzymes ʻelua he ʻano like loa, a he ʻano like nō hoʻi ka hui amino acid, akā ʻokoʻa ka helu o ka aspartic acid a me nā koena proline.ʻO Guangxi king cobra snake venom ka waiwai PLA2.Shu Yuyan et al.(1989) hoʻokaʻawale i kahi PLA2 mai ka mea make, he 3.6 mau manawa kiʻekiʻe ma mua o ka lāʻau make mua, kahi paona molekala o 13000, kahi hui o 122 amino acid koena, kahi isoelectric point o 8.9, a me ke kūpaʻa wela maikaʻi.Mai ka electron microscope ka nānā ʻana i ka hopena o ka PLA2 kumu ma luna o nā keʻena koko ʻulaʻula, hiki ke ʻike ʻia he hopena ʻike ʻia i ka membrane o ke koko ʻulaʻula kanaka, akā ʻaʻohe hopena i ʻike ʻia i nā kao ʻulaʻula ʻulaʻula.Loaʻa i kēia PLA2 ka hopena hoʻopaneʻe i ka wikiwiki electrophoretic o nā keʻena koko ʻulaʻula i loko o ke kanaka, nā kao, nā lapeti a me nā puaʻa guinea.ʻO Chen et al.Hiki i kēia enzyme ke pale i ka hōʻuluʻulu platelet i hoʻokomo ʻia e ADP, collagen a me ka sodium arachidonic acid.Ke 10/xg/ml~lOOjug/ml ka nui o ka PLA2, ua pale loa ia ka houluulu platelet.Inā hoʻohana ʻia nā platelets i holoi ʻia e like me nā mea, ʻaʻole hiki i ka PLA2 ke kāohi i ka hoʻohui ʻana ma ke ʻano o 20Mg/ml.ʻO Aspirin kahi mea pale o cyclooxygenase, hiki ke pale i ka hopena o PLA2 ma nā platelets.Hiki i ka PLA2 ke keʻakeʻa i ka hōʻuluʻulu platelet ma ka hydrolyzing arachidonic acid e synthesize thromboxane A2.Ua aʻo ʻia ka conformation solution o PLA2 e Agkistrodon halys Pallas venom ma Zhejiang Province ma o ka circular dichroism, fluorescence a me UV absorption.Ua hōʻike ʻia nā hopena hoʻokolohua ua like ke ʻano o ke kaulahao nui o kēia enzyme me ke ʻano like o ka enzyme mai nā ʻano ʻē aʻe a me nā genera, ʻoi aku ka maikaʻi o ka hoʻohālikelike ʻana i ka wela, a ua hiki ke hoʻololi ʻia ka hoʻololi ʻana o ka hoʻololi ʻana i ke kaiapuni waikawa.ʻAʻole pili ka hui pū ʻana o ka activator Ca2 + a me ka enzyme i ke kaiapuni o nā koena tryptophan, ʻoiai ʻo Zn2 + inhibitor e hana ʻē aʻe.ʻOkoʻa ke ala e pili ai ka waiwai pH o ka hopena i ka hana enzyme mai nā reagents i luna.
I ke kaʻina hana o ka PLA2 hoʻomaʻemaʻe i ka ʻawaʻawa o ka nahesa, he mea ʻike maopopo ʻia ʻo ka nahesa he ʻelua a ʻoi aʻe paha nā piko elution PLA2.Hiki ke wehewehe ʻia kēia hanana penei: ① ma muli o ke ola ʻana o nā isozymes;② Hoʻokahi ʻano PLA2 i polymerized i loko o nā ʻano hui like ʻole PLA2 me nā paona molekala like ʻole, ʻo ka hapa nui o ia mau mea ma ka laulā o 9 000 ~ 40 000;③ ʻO ka hui pū ʻana o PLA2 a me nā mea ʻawaʻawa snake ʻē aʻe e paʻakikī iā PLA2;④ No ka hydrolyzed ka pilina amide ma PLA2, hoʻololi ka uku.① A he mea maʻamau ka ②, me nā ʻokoʻa liʻiliʻi wale nō, e like me PLA2 ma CrWa/w snake venom
ʻElua kūlana: ① a me ②.Ua ʻike ʻia ke kolu o ka maʻi ma PLA2 i ka mea make o nā nahesa penei: Oxyranus scutellatus, Parademansia microlepidota, Bothrops a ^>er, Palestinian viper, sand viper, a me rattlesnake weliweli km.
ʻO ka hopena o ka hihia ④ e hoʻololi i ka wikiwiki o ka neʻe ʻana o PLA2 i ka wā electrophoresis, akā ʻaʻole e loli ka hui ʻana o ka amino acid.Hiki ke wāwahi ʻia nā peptide e ka hydrolysis, akā naʻe, paʻa mau lākou e nā paʻa disulfide.Aia i loko o ka ʻawaʻawa o ka lua hikina nā ʻano PLA2, i kapa ʻia ʻo type a a me ka type p PLA2.ʻO ka ʻokoʻa ma waena o kēia mau ʻano ʻelua o PLA2 he hoʻokahi amino acid, ʻo ia hoʻi, hoʻololi ʻia ka glutamine i hoʻokahi mole PLA2 e ka glutamic acid i ka mole PLA2 ʻē aʻe.ʻOiai ʻaʻole maopopo ke kumu kūpono o kēia ʻokoʻa, ua manaʻo nui ʻia e pili ana i ka deamination o PLA2.Inā mālama ʻia ka PLA2 i loko o ka viper viper Palestinian me ka mea ʻawaʻawa, e lilo nā pūʻulu hope i loko o kāna mau molekele enzyme ma mua o ka wā ma mua.Mai ka C PLA2 i hoʻokaʻawale ʻia mai ka nahesa ʻawaʻawa ʻelua ʻokoʻa N-terminal, a ʻo kona kaumaha molekala he 30000. Hiki ke kumu kēia ʻano ma muli o ka dimer asymmetric o PLA2, e like me ka dimer symmetric i hoʻokumu ʻia e PLA2 i ka mea make o ka nahesa diamondback rattlesnake hikina. a me ka nahesa rattlesnake o ke komohana.He nui nā ʻano subspecies ka ʻĀkia cobra, ʻaʻole maopopo loa kekahi o ia mau mea i ka hoʻohālikelike ʻana.No ka laʻana, ʻike ʻia ka mea i kapa ʻia ʻo Cobra Outer Caspian subspecies i kēia manawa
Pono ia e pili i ka Outer Caspian Sea Cobra.No ka mea he nui nā ʻano subspecies a hui pū ʻia, ʻokoʻa ke ʻano o ka snake venom ma muli o nā kumu like ʻole, a kiʻekiʻe hoʻi ka ʻike o nā isozymes PLA2.No ka laʻana, ka make ʻawaʻawa
Ma kahi o 9 mau ʻano isozymes PLA2 o nā ʻano like ʻole i loaʻa i loko, a ʻo 7 mau ʻano isozymes PLA2 i loaʻa i loko o ka mea ʻawaʻawa o nā subspecies Caspian.ʻO Durkin et al.(1981) ua aʻo i ka PLA2 maʻiʻo a me ka helu o nā isozymes i nā ʻawaʻawa nahesa like ʻole, me 18 mau ʻawaʻawa cobra, 3 mau mea ʻawaʻawa mamba, 5 mau ʻawaʻawa ʻawaʻawa, 16 mau ʻawaʻawa rattlesnake a me 3 mau mea ʻawaʻawa moana.Ma keʻano laulā, kiʻekiʻe ka hana PLA2 o ka cobra cobra, me nā isozymes he nui.ʻO ka hana PLA2 a me nā isozymes o ka viper venom he medium.He haʻahaʻa loa ka hana PLA2 o ka mamba venom a me rattlesnake a i ʻole ka hana PLA2.He haʻahaʻa hoʻi ka hana PLA2 o ka nahesa moana.
I nā makahiki i hala iho nei, ʻaʻole i hōʻike ʻia aia ka PLA2 i loko o ka snake venom ma ke ʻano o ka dimer ikaika, e like me ka hikina rhombophora rattlesnake (C. , a ʻo ka dimerase wale nō
Ka hana.Shen et al.ʻO ka dimer wale nō o ka PLA2 o ka mea make nahesa ke ʻano ikaika o ka enzyme.ʻO ke aʻo ʻana i ka hoʻolālā spatial e hōʻike pū ana aia ka PLA2 o ke komohana diamondback rattlesnake ma ke ʻano o ka dimer.Pūhui piscivorous
Elua okoa PLA ^ Ei a me E2 o ka make nahesa, aia 仏 ma ke ano o ka dimer, he hana ka dimer, a he hana ole kona monomer i hookukeia.Lu Yinghua et al.(1980) aʻo hou i ke kino a me ke kemika waiwai a me ka pane kinetics o E. Jayanthi et al.(1989) hoʻokaʻawale i kahi PLA2 kumu (VRVPL-V) mai ka viper viper.ʻO ke kaumaha molekala o ka monomer PLA2 he 10000, nona ka hopena lethal, anticoagulant a me ka edema.Hiki i ka enzyme ke polymerize i nā polymers me nā paona molekala like ʻole ma lalo o ke kūlana o PH 4.8, a hoʻonui ke degere o ka polymerization a me ke kaumaha o nā polymers me ka piʻi ʻana o ka mahana.ʻO ke kaumaha molekala o ka polymer i hana ʻia ma 96 ° C ʻo 53 100, a hoʻonui ka hana PLA2 o kēia polymer i ʻelua.
Ka manawa hoʻouna: Nov-18-2022