Sunta abeesadu waxay ka kooban tahay ensaymes kuwaas oo hydrolyzed kaarboxyl ester bonds.Substrates for hydrolysis waa phospholipids, acetylcholine iyo acetate aromatic.Enzymes-yadaas waxaa ka mid ah saddex nooc: phospholipase, acetylcholinesterase iyo aromatic esterase.Arginine esterase ee sunta abeesadu waxay sidoo kale hydrolyze kartaa arginine synthetic ama lysine, laakiin inta badan waxay hydrolyzed bond bonds peptide ee dabiiciga ah, sidaas darteed waxaa iska leh protease.Enzymes-yada halkan lagaga hadlay waxay ku shaqeeyaan oo keliya substrates-ka ester-ka mana ku dhaqmi karaan curaarta peptide kasta.Waxaa ka mid ah enzymes-ka, hawlaha bayooloji ee acetylcholinesterase iyo phospholipase ayaa aad uga muhiimsan oo si buuxda loo bartay.Qaar ka mid ah sunta abeesada waxay leeyihiin firfircooni udgoon udgoon, kaas oo hydrolyze kara p-nitrophenyl ethyl ester, a - ama P-naphthalene acetate iyo indole ethyl ester.Ilaa hadda lama garanayo in hawshan ay soo saartay ensaymes madaxbannaan ama saameyn la og yahay ee karboksylesterase, iska daa muhiimadeeda noole.Marka sunta Agkistrodon halys Japonicus lagu falceliyay p-nitrophenyl ethyl ester iyo indole ethyl ester, hydrolysates ee p-nitrophenol iyo indole phenol lama helin;Taa beddelkeeda, haddii esters-yadani ay ka falceliyaan noocyada cobra Zhoushan ee maska iyo sunta maska ee Bungarus multicinctus, si dhakhso ah ayaa loo nadiifin doonaa.Waxaa la og yahay in suntan cobra ay leeyihiin firfircoonida cholinesterase xoog leh, taas oo laga yaabo inay mas'uul ka tahay hydrolysis ee substrates kor ku xusan.Xaqiiqdii, Mclean et al.(1971) ayaa sheegay in sun badan oo masaska ah oo ka tirsan qoyska Cobra ay hydrolyze karaan indole ethyl ester, naphthalene ethyl ester iyo butyl naphthalene ester.Suntan masaska ah waxay ka kala yimaaddaan: kobra, qoor madow, kobra dibnaha madow, kobra dahabka, masar masar, kobra boqor, mamba mamba, mamba madow iyo mamba dibinta cad (D. aw weli wuu yaqaan rhombola rattlesnake bari
Sunta abeesadu waxay hydrolyze kartaa methyl indole ethyl ester, kaas oo ah substrate-ka lagu go'aamiyo dhaqdhaqaaqa cholinesterase ee serum, laakiin suntan maska ahi ma muujinayso dhaqdhaqaaqa cholinesterase.Tani waxay tusinaysaa in uu jiro emsterase aan la garanayn oo ku jira sunta cobra, taas oo ka duwan cholinesterase.Si loo fahmo dabeecadda ensaymeskan, shaqo dheeraad ah oo kala soocid ayaa loo baahan yahay.
1. Phospholipase A2
(I) Dulmar
Phospholipase waa enzim ka saari kara phosphate glyceryl.Waxaa jira 5 nooc oo phospholipase ah oo dabiiciga ah, kuwaas oo kala ah phospholipase A2 iyo phospholipase
A., phospholipase B, phospholipase C iyo phospholipase D. Sunta masku waxay inta badan ka kooban tahay phospholipase A2 (PLA2), sunta abeesada dhowr ah ayaa ka kooban phospholipase B, fosfolipases kale ayaa inta badan laga helaa unugyada xayawaanka iyo bakteeriyada.Jaantuska 3-11-4 waxa uu tusayaa goobta falgalka ee fosfoolipases-ka ee ku saabsan hydrolysis substrate.
Ka mid ah fosfoolipases, PLA2 ayaa wax badan laga bartay.Waxa laga yaabaa inay tahay enzyme-ka ugu badan ee sunta abeesada ku jirta.Substrate-keedu waa dammaanadda ester-ka ee booska labaad ee Sn-3-glycerophosphate.Insaymamkan waxa si weyn looga helaa sunta abeesada, sunta shinnida, sunta Dabaqaalaha iyo unugyada xoolaha, PLA2 waxa ay aad ugu badan tahay afar qoys oo suntan masaska ah.Sababtoo ah enzyme-kan ayaa jebiya unugyada dhiigga cas wuxuuna keenaa hemolysis, waxaa sidoo kale loo yaqaan "hemolysin".Dadka qaar ayaa sidoo kale u yaqaan PLA2 hemolytic lecithinase.
Ludeeke ayaa markii ugu horeysay ogaatay in sunta abeesadu ay soo saari karto iskudhis hemolytic ah oo ku shaqeeya lecithin iyada oo loo marayo enzymes.Later, Delezenne et al.Waxay caddeeyeen in marka sunta cobra ay ka shaqeyso serum faras ama jaallo ah, ay sameyso walax hemolytic ah.Hadda waxaa la og yahay in PLA2 ay si toos ah ugu dhaqmi karto fosfoolipids ee xuubka erythrocyte, burburinta qaabdhismeedka xuubka erythrocyte oo keena dhiig-bax toos ah;Waxa kale oo ay ku dhaqmi kartaa serum ama lecithin lagu daray si ay u soo saarto lecithin hemolytic, kaas oo ka shaqeeya unugyada dhiigga cas si ay u soo saaraan hemolysis aan toos ahayn.Inkasta oo PLA2 ay ku badan tahay afarta qoys ee sunta abeesada, waxa ku jira enzymes-ka suntan abeesooyinka kala duwan ayaa xoogaa ka duwan.Rattlesnake (C
Sunta abeesada waxa ay muujisay firfircooni PLA2 liidata.Shaxda 3-11-11 waxay muujineysaa isbarbardhigga waxqabadka PLA2 ee 10 sunta waaweyn ee masaska sunta ah ee Shiinaha.
Shaxda 3-11-11 Isbarbardhigga hawlaha phospholipase VIII ee 10 sunta masaska ee Shiinaha
Sunta abeesada
Dufanka sii daaya
Aliphatic acid,
Cjumol/mg)
Dhaqdhaqaaqa hemolytic CHU50/^ g * ml)
Sunta abeesada
Sii daa asiidhyada dufanka leh
(^raol/mg)
Dhaqdhaqaaqa hemolytic "(HU50/ftg * 1111)
Najanaja atra
9.62
kow iyo toban
Microcephal ophis
shan dhibic kow eber
kalyspallas
8.68
laba kun iyo siddeed boqol
gracilis
V, acutus
7.56
* * #
Ophiophagus hannah
saddex dhibic siddeed iyo laba
boqol iyo afartan
Bnugarus faktatus
7,56
laba boqol iyo siddeetan
B. multicinctus
hal dhibic sagaal lix
laba boqol iyo siddeetan
Viper iyo russelli
toddoba dhibcood eber saddex
T, mukrosquamatus
hal dhibic sideed iyo shan
Siamensis
T. stejnegeri
0. 97
(2) Kala saarida iyo nadiifinta
Waxa ku jira PLA2 ee sunta abeesada way weyn tahay, wayna degan tahay kulaylka, aashitada, alkali iyo denaturant, si ay u fududaato in la nadiifiyo oo la kala saaro PLA2.Habka caadiga ah waa in marka hore la sameeyo sifeynta jel ee sunta cayriin, ka dibna la sameeyo chromatography beddelashada ion, tallaabada xigtana waa la soo celin karaa.Waa in la ogaadaa in qaboojinta-qallajinta ee PLA2 ka dib ion-exchange chromatography waa inaysan keenin isugeyn, sababtoo ah habka qalajinta qalajinta inta badan waxay kordhisaa xoogga ionic ee nidaamka, taas oo ah arrin muhiim ah oo keenaysa isku-darka PLA2.Marka lagu daro hababka guud ee kor ku xusan, hababka soo socda ayaa sidoo kale la qaatay: ① Wells et al.Analoogga substrate-ka ee PLA2 ayaa loo isticmaalay sidii ligand ee chromatography-ga xidhiidhka.Liinkan waxa uu ku xidhi karaa PLA2 sunta masaska oo leh Ca2+.EDTA waxaa inta badan loo isticmaalaa sidii wax-soo-saarka.Ka dib markii Ca2+ meesha laga saaro, xidhiidhka u dhexeeya PLA2 iyo ligand wuu yaraanayaa, waxaana laga go'i karaa ligand.Kuwo kale waxay isticmaalaan 30% xal organic ama 6mol/L urea sida eluent.③ Hydrophobic chromatography waxaa lagu sameeyay PheiiylSephar0SeCL-4B si meesha looga saaro raadadka PLA2 ee ku jira cardiotoxin.④ Antibody-ka PLA2 waxaa loo isticmaalay sidii ligand si loogu sameeyo chromatography affinity on PLA2.
Ilaa hadda, tiro badan oo sunta masaska ah ee PLAZ ayaa la nadiifiyey.Tu iyo al.(1977) waxay taxday PLA2 oo laga sifeeyay sunta abeesada ka hor 1975. Sannadihii u dambeeyay, tiro badan oo maqaallo ah oo ku saabsan kala-soocidda iyo nadiifinta PLA2 ayaa la soo sheegay sannad kasta.Halkan, waxaan diiradda saareynaa kala-soocidda iyo nadiifinta PLA ee culimada Shiinaha.
Chen Yuancong iyo al.(1981) waxay kala sooceen saddex nooc oo PLA2 ah oo ka soo jeeda sunta Agkistrodon halys Pallas ee Zhejiang, kuwaas oo loo qaybin karo acidic, dhexdhexaad ah iyo alkaline PLA2 marka loo eego dhibcaha isoelectric.Marka loo eego sunta, PLA2 dhexdhexaad ah ayaa aad u sun badan, kaas oo loo aqoonsaday inuu yahay neurotoxin presynaptic Agkistrodotoxin.Alkaline PLA2 way ka yar tahay sunta, iyo PLA2 aashitada leh ku dhawaad wax sun ah ma laha.Wu Xiangfu iyo al.(1984) marka la barbardhigo sifooyinka saddexda PLA2s, oo ay ku jiraan miisaanka kelli, halabuurka amino acid, N-terminal, barta isoelectric, xasilloonida kuleylka, dhaqdhaqaaqa enzyme, sunta iyo dhaqdhaqaaqa hemolytic.Natiijooyinku waxay muujiyeen inay leeyihiin miisaan isku mid ah iyo xasilloonida kulaylka, laakiin waxay ku kala duwan yihiin dhinacyo kale.Marka la eego dhaqdhaqaaqa enzyme, waxqabadka enzyme acid wuxuu ka sarreeyaa waxqabadka enzyme alkaline;Saamaynta hemolytic ee enzyme alkaline ee unugyada dhiigga cas ee jiirka ayaa ahaa kan ugu xoogga badan, oo ay ku xigto ensaym dhexdhexaad ah, iyo ensaymka aashitada oo aan si dhib leh u dhiigxin.Sidaa darteed, waxaa la qiyaasayaa in saameynta hemolytic ee PLAZ ay la xiriirto kharashka PLA2 molecule.Zhang Jingkang iyo al.(1981) waxay sameeyeen Agkistrodotoxin crystals.Tu Guangliang iyo al.(1983) ayaa sheegay in PLA sun ah oo leh barta isoelectric ee 7. 6 waa la go'doomiyay oo laga nadiifiyay sunta Vipera rotundus ee Fujian, iyo sifooyinka jireed iyo kiimiko, halabuurka amino acid iyo taxanaha 22 amino acid haraaga N. - terminal ayaa la go'aamiyay.Li Yuesheng iyo al.(1985) ayaa ka go'ay oo ka nadiifiyay PLA2 kale sunta Viper rotundus ee Fujian.Qeybta hoose ee PLA2 * waa 13 800, barta isoelectric waa 10.4, waxqabadka gaarka ah waa 35/xnioI/miri mg. LD5 ayaa xididka laga duray jiirarka.Waa 0.5 ± 0.12mg/kg.Insaymiskani wuxuu leeyahay saameyn cad oo xinjirowga lidka ku ah iyo hemolytic.Unugyada sunta ah ee PLA2 waxa ay ka kooban yihiin 123 hadhaa ah oo ah 18 nooc oo amino acids ah.Moleekulku wuxuu qani ku yahay cysteine (14), aspartic acid (14) iyo glycine (12), laakiin kaliya waxa ku jira hal methionine, N-terminal-kiisuna waa serine hadhaa.Marka la barbar dhigo PLA2 ee ay go'doomisay Tuguang, miisaanka kelli iyo tirada hadhaaga amino acid ee labada isoenzymes aad bay isugu shabahaan, iyo ka koobanaanta amino acid sidoo kale aad bay isugu dhow yihiin, laakiin tirada aspartic acid iyo hadhaaga proline way ka duwan yihiin.Sunta maska cobra ee boqorka Guangxi waxay ka kooban tahay PLA2 qani ah.Shu Yuyaan iyo al.(1989) ayaa PLA2 ka soocay sunta, taas oo leh waxqabad gaar ah 3.6 jeer ka sarreeya sunta asalka ah, miisaanka kelli ee 13000, ka kooban 122 amino acid haraaga, barta isoelectric ee 8.9, iyo xasilloonida kulaylka wanaagsan.Marka la eego mikroskoob elektaroonig ah oo ku saabsan saamaynta PLA2 aasaasiga ah ee unugyada dhiigga cas, waxaa la arki karaa in ay saameyn muuqata ku leedahay xuubabka unugga dhiigga cas ee bini'aadamka, laakiin wax saameyn ah kuma laha unugyada cas cas ee riyaha.PLA2-kan waxa uu leeyahay saamayn dibudhac cad oo ku saabsan xawaaraha korantada ee unugyada dhiigga cas ee dadka, riyaha, bakaylaha iyo doofaarrada guinea.Chen iyo al.Insaymiskani waxa uu hor istaagi karaa isku-darka platelet-ka oo ay keentay ADP, collagen iyo sodium arachidonic acid.Marka fiirsashada PLA2 ay tahay 10/xg/ml~lOOjug/ml, isu geynta platelet gabi ahaanba waa la joojinayaa.Haddii platelets-ka la dhaqay loo isticmaalo walxo ahaan, PLA2 ma joojin karto isku-darka 20Mg/ml.Aspirin waa xannibaadaha cyclooxygenase, kaas oo joojin kara saameynta PLA2 ee platelets.PLA2 waxaa laga yaabaa inay joojiso isku-darka platelet-ka iyadoo hydrolyzing arachidonic acid si ay u abuurto thromboxane A2.Xalka qaabaynta PLA2 ee ay soo saartay Agkistrodon halys Pallas venom ee Gobolka Zhejiang waxa lagu bartay iyada oo la adeegsanayo dichroism wareeg ah, fluorescence iyo nuugista UV.Natiijooyinka tijaabada ah waxay muujiyeen in isku-dhafka silsiladda ugu weyn ee ensaymkani uu la mid yahay midka isku midka ah ee enzym ka noocyada kale ee noocyada kale, qaabka qalfoofka ayaa lahaa caabbinta kulaylka wanaagsan, iyo isbeddelka qaabdhismeedka jawiga aashitada ayaa dib loo rogay.Isku darka activator Ca2+ iyo enzim ma saameeyaan deegaanka haraaga tryptophan, halka inhibitor Zn2+ uu sameeyo lidkeeda.Habka uu qiimaha pH ee xalku u saameeyo dhaqdhaqaaqa enzyme wuu ka duwan yahay reagen-yaasha sare.
Habka nadiifinta PLA2 ee sunta abeesada, ifafaale muuqda ayaa ah in sunta abeesada ay ka kooban tahay laba ama in ka badan oo PLA2 elution ah.Dhacdadan waxaa lagu sharxi karaa sida soo socota: ① sababtoo ah jiritaanka isozymes;② Hal nooc oo PLA2 ah ayaa lagu dhejiyay noocyo kala duwan oo PLA2 ah oo isku dhafan oo leh miisaanno unugyo kala duwan, kuwaas oo intooda badan ku jira 9 000 ~ 40 000;③ Isku darka PLA2 iyo qaybaha kale ee sunta abeesada ayaa adkeynaysa PLA2;④ Sababtoo ah curaarta amide ee PLA2 waa hydrolyzed, kharashka ayaa isbedelaya.① Iyo ② waa wax caadi ah, marka laga reebo dhowr ka reeban, sida PLA2 ee sunta maska ee CrWa/w
Waxaa jira laba xaaladood: ① iyo ②.Xaaladda saddexaad ayaa laga helay PLA2 ee sunta masaska soo socda: Oxyranus scutellatus, Parademansia microlepidota, Bothrops a ^>er, Viper Falastiin, Viper ciid, iyo rattlesnake km.
Natiijada kiiska ④ waxay ka dhigaysaa xawaaraha socdaalka PLA2 isbeddelka inta lagu jiro electrophoresis, laakiin ka kooban amino acid isma beddelo.Peptides waxaa lagu jebin karaa hydrolysis, laakiin guud ahaan waxay weli ku xiran yihiin curaarta disulfide.Sunta godka bari ee rattlesnake waxay ka kooban tahay laba nooc oo PLA2 ah, oo loo yaqaan nooca a iyo nooca p PLA2 siday u kala horreeyaan.Farqiga u dhexeeya labadan nooc ee PLA2 waa hal amino acid oo keliya, taas oo ah, glutamine ee ku jira hal unug oo PLA2 ah ayaa lagu beddelay glutamic acid ee molecule kale ee PLA2.Inkasta oo sababta dhabta ah ee kala duwanaanshuhu aanay caddayn, waxaa guud ahaan la rumeysan yahay inay la xiriirto deamination-ka PLA2.Haddii PLA2 ee sunta viper ee falastiiniyiinta lagu hayo diirimaad sunta cayriin, kooxaha dhammaadka ah ee unugyadeeda ensaymku waxay noqonayaan wax ka badan sidii hore.Laga soo bilaabo C PLA2 oo ka go'doomay sunta abeesadu waxay leedahay laba N-terminal oo kala duwan, miisaankeeduna unugyadu waa 30000. Dhacdadan waxaa sababi kara dimer-ka asymmetric ee PLA2, kaas oo la mid ah dimer-ka sumaysan ee ay samaysay PLA2 ee sunta bari ee dheemanback rattlesnake. iyo reer galbeedka diamondback rattlesnake.Cobra Aasiya waxay ka kooban tahay noocyo badan oo kala duwan, kuwaas oo qaarkood aan si qeexan loo kala saarin.Tusaale ahaan, waxa la odhan jiray Cobra Outer Caspian-hoosaadka hadda waa la aqoonsaday
Waa in loo nisbeeyaa Badda Caspian-ka ee Cobra.Maadaama ay jiraan noocyo badan oo kala duwan oo ay isku dhafan yihiin, isku dhafka sunta abeesada aad ayey u kala duwan tahay ilo kala duwan awgeed, waxa ku jira PLA2 isozymes sidoo kale waa sarreeyaa.Tusaale ahaan, sunta cobra
Ugu yaraan 9 nooc oo PLA2 isozymes ah oo noocyada r ll ah ayaa laga helay, iyo 7 nooc oo PLA2 isozymes ah ayaa laga helay sunta noocyada cobra ee Caspian.Durkin iyo al.(1981) waxa uu daraaseeyay nuxurka PLA2 iyo tirada isozymes ee ku jira sunta abeesooyinka kala duwan, oo ay ku jiraan 18 sunta cobra, 3 suntan mamba, 5 sunta viper, 16 suntan masska ah iyo 3 suntan masaska badda.Guud ahaan, waxqabadka PLA2 ee sunta cobra waa mid sarreeya, oo leh isozymes badan.Hawlaha PLA2 iyo isozymes ee sunta viper-ka waa dhexdhexaad.Dhaqdhaqaaqa PLA2 ee sunta mamba iyo sunta masska waa mid aad u hooseeya ama ma jiro hawl PLA2 ah.Dhaqdhaqaaqa PLA2 ee sunta masaska badda ayaa sidoo kale hooseysa.
Sannadihii u dambeeyay, lama soo sheegin in PLA2 ee sunta abeesadu ay u jirto qaab dimer firfircoon, sida rhombophora rattlesnake bari (C. sunta masku waxay ka kooban tahay nooca a iyo nooca PLA2, labaduba waxay ka kooban yihiin laba qaybood oo isku mid ah. , oo kaliya dimerase ayaa leh
DhaqdhaqaaqaShen iyo al.Waxa kale oo la soo jeediyay in kaliya dimer PLA2 ee sunta abeesada ay tahay qaabka firfircoon ee enzyme.Daraasadda qaab-dhismeedka bannaan waxay sidoo kale caddaynaysaa in PLA2 ee rattlesnake dheeman dhabarka galbeed uu u jiro qaab dimer ah.Iskudhis Piscivorous
Waxaa jira laba kala duwan oo PLA ^ Ei iyo E2 ee sunta abeesada, kuwaas oo 仏 ku jira qaabka dimer, dimer-ku waa firfircoon yahay, monomer-ka kala go'a waa mid aan firfircoonayn.Lu Yinghua iyo al.(1980) wuxuu sii bartay sifooyinka jirka iyo kiimikaad iyo kinetics falcelinta E. Jayanthi et al.(1989) waxay ka go'day PLA2 aasaasiga ah (VRVPL-V) ee sunta viper.Miisaanka molecular ee monomer PLA2 waa 10000, kaas oo leh saameyn dilaa ah, xinjirowga lidka ku ah iyo bararka.Insaymku wuxuu polymerize karaa polymers leh miisaanyo kala duwan oo kelli ah oo hoos yimaada xaaladda PH 4.8, iyo heerka polymerization iyo miisaanka kelli ee polymers-ka ayaa kordhiya kororka heerkulka.Miisaanka molecular ee polymer-ka laga dhaliyo 96 ° C waa 53 100, iyo waxqabadka PLA2 ee polymerkani wuxuu kordhiyaa laba
Waqtiga boostada: Nov-18-2022