Ksantoproteinovaya asabela ngayo amaprotheni: izibonakaliso ifomula kwesibalo

Ukuze izinhlangano zikwazi ukusungula ukwakheka qualitative ukudla abaningi babesebenzisa ksantoproteinovaya yokusabela kuya amaprotheni. Ukuba khona acid elimnandi acid e ukuxubana zizoshintsha ngendlela eyakhayo test isampula umbala.

Kuyini amaprotheni

Ibizwa nangokuthi iprotheni, okuyinto yokwakha ingeyomzimba eziphilayo. Amaphrotheni silondoloze usayizi imisipha, ukubuyisela elimele abese efile izicubu izakhiwo kwezitho ahlukahlukene, kungaba izinwele, isikhumba kanye nemisipha. With iqhaza zabo, ukhiqiza abomvu corpuscles, ilawulwa ukuhlinzwa evamile yama-hormone amaningi omzimba amaseli.

Lena molecule eyinkimbinkimbi ukuthi kuyinto polypeptide kokuba mass ngaphezu kuka 6 ~ 10 ³ daltons. Amaphrotheni isakhiwo ukwakha acid acid izinsalela e inqwaba oluhambisana peptide isibopho.

Isakhiwo amaphrotheni

Isici esiphawulekayo lezi izinto uma kuqhathaniswa ne ongaphakeme yamangqamuzana isisindo peptides wukuthi bajwayela isakhiwo ngakuthathu kwendawo, asekelwe imiphumela nge ntshashintshe okudonsa. Amaphrotheni ifa emine isakhiwo. Ukuze ngamunye wabo has izici zayo siqu.

Njengoba isisekelo lenhlangano eyinhloko yezinhlayiya etholakala acid acid ukulandelana, isakhiwo esingagcina ibona ksantoproteinovaya yokusabela kuya amaprotheni. Lesi sakhiwo kuyinto ngezikhathi ngokuphinda peptide isibopho -HN-CH-ngokubambisana, a abakhethayo ingxenye ezizivelelayo kukhona amaketanga uhlangothi amino acid. Abakuchaza izakhiwo eminye efana iyonke.

Okuxhunyanwa amaprotheni isakhiwo kubhekwa uqine ngokwanele, lokhu kungenxa yokuba khona ekuxhumaneni eqinile covalent e peptide izibopho. Ukwakheka amazinga ezalandela kwenzeka kuye izimfanelo isethwe ku esigabeni sokuqala.

Ukwakheka isakhiwo yesibili kungenzeka ngenxa kuhlanekezela ukulandelana acid acid in a Kuvunguza, lapho i-hydrogen izibopho amiswa phakathi ematfuba.

Zemfundo ephakeme ezingeni inhlangano zinhlayiya zakha ngokusebenzisa omunye Kuvunguza ku nezinye izindengezi nge okuvela uxhumano kungenzeka phakathi kwabo, ne-hydrogen, disulfide, covalent noma kwakwakhe ionic. Umphumela inhlangano efomini globules.

Leli lungiselelo kwezendawo izakhiwo ephakeme lapho kumiswa izibopho zamakhemikhali phakathi kwawo kuholela ekwakhekeni yefomu yokugcina engqamuzaneni noma amazinga quaternary.

amino acid

Enza nezakhi ezithile zamakhemikhali amaprotheni. Kukhona acid ezinkulu acid angu-20 abakha polypeptides ilandelane ezahlukene. Lokhu kubizwa ngokuthi ezingavamile aminocarboxylic acid hydroxyproline ifomu hydroxylysine etholakala peptide eziyisisekelo.

Ngenxa uphawu ukuqashelwa ksantoproteinovaya amaprotheni ukuphendula, khona acid ezithile acid inikeza ushintsho umbala reagents, esikisela khona izinhlaka eziqondene Ukwakheka kwabo.

Kwabe sekuvela ukuthi, zingabantu acid carboxylic ezenzeke esikhundleni hydrogen-athomu iqembu acid.

Kuvele isakhiwo Ngokwesibonelo engqamuzaneni zingaba a kwesakhiwo ifomula glycine (HNH- HCH- COOH) njengoba elula acid acid.

Kulokhu omunye hydrogens CH ₂ - kungenzeka indawo carbon eside olukhulu ehlanganisa i-benzene indandatho, acid, sulfo, carboxy.

Yini ksantoproteinovaya ukuphendula

Ukuze ukuhlaziywa qualitative amaprotheni besebenzisa amasu ahlukahlukene. Lezi zihlanganisa ukusabela:

• biuret nge ukubukeka umbala violet;
• ninhydrin ukwakha isixazululo blue-violet;
• formaldehyde ne ukusungulwa umbala obomvu;
• Nogier ukuba kwenze umbala grey-black.

Lapho efeza indlela ngayinye kwafakazela khona amaprotheni khona amaqembu athile ukusebenza i-molecule yabo.

Kube ksantoproteinovaya yokusabela kuya amaprotheni. Ibizwa nangokuthi ukuwohloka Mulder. Libhekisela ukusabela umbala amaprotheni, lapho kukhona enamakha heterocyclic amino acid.

Isici esivelele sesimiso inqubo isampula ezifana kuyinto nitration acid nitric eyindilinga acid acid izinsalela ngokukhethekile ukubusa leqembu nitro iringi benzene.

Waba yini umphumela walesi nqubo kumiswa nitro compounds, okuyinto ukugqashuka. Lona eziyisisekelo sici ksantoproteinovaya ukuphendula.

Yini ezanquma acid acid

Akubona bonke aminocarboxylic acid kungatholakala usebenzisa isampula enjalo. Isici esiyinhloko ksantoproteinovaya amaprotheni ukuthola yokusabela - khona iringi benzene noma indandatho heterocyclic e acid-molecule acid.

Ngoba namaprotheni behlukene acid aminocarboxylic ezimbili elimnandi, lapho kukhona iqembu phenyl (I-phenylalanine), kanye hydroxyphenyl olukhulu (e tyrosine).

Nge ksantoproteinovaya ukusabela kunqunywa heterocyclic acid acid tryptophan, indole nokuba nucleus elimnandi. Ukuba khona zezakhi ngenhla amaprotheni kunikeza isici umbala ushintsho kwemvelo yokuhlola.

Yini ukusetshenziswa reagents

Sokufeza ukusabela ksantoproteinovaya kudingeka ukulungiselela isixazululo 1% Iqanda amaprotheni, noma imvelaphi zemifino.

Ngokuvamile esetshenziswa amaqanda, okuyinto ihlukaniswe ngokwahlukana okunye amaprotheni kusukela isikhuphasha. Ukuze uthole isixazululo 1% of amaprotheni ukuthi buhlanjululwe in a inani ngokuphindwe kayishumi yamanzi hlamba. Ngemva kokuphela okuholela amaprotheni ketshezi okufanele kuhlungwe ngokusebenzisa izingqimba eziningana cheesecloth. Lokhu Isixazululo kufanele agcinwe endaweni epholile.

Kungenzeka ukuba sifeze ukuphendula iprotheni zemifino. Ukulungisa isixazululo isetshenziswa ufulawa kakolweni e isamba 0.04 kg. 0,16 l yamanzi hlamba. Izithako abangu kuxutshwe negabha okuyinto kuboshelwe amahora 24 endaweni epholile ezingeni lokushisa mayelana + 1 ° C. Ekupheleni kosuku ikhambi kwakudalwa, kulandele filtration yalo yokuqala nge uhala bese - isisefo sephepha eligoqiwe. Lolu ketshezi wafakwa igcinwe endaweni epholile. Lokhu Isixazululo ikakhulu ekhona ingxenyenamba-albumin.

Ukuze ofeza ksantoproteinovaya ukuphendula njengoba main reagent esetshenziswa nitric acid Conc. Ngaphezu kwalokho reagents yisisombululo of 10% sodium hydroxide noma ammonia, ikhambi gelatin futhi phenol unconcentrated.

Le ndlela yokwenza we

Ngo ngamapayipi ehlanzekile senziwa 1% isixazululo amaprotheni amaqanda -th noma kafulawa isamba 2 ml. Ukuze lokhu wanezela mayelana 9 amathonsi kanzulu nitric acid ukuze flocculation kumisiwe. Ingxube wafakwa ukufuthelana, okuholela ophuzi kwenze kancane kancane ubalekela futhi umbala waso ingena isixazululo.

Lapho ketshezi epholisa phansi tube ngabambelela odongeni ngihamba wanezela mayelana 9 amathonsi sodium hydroxide, bagxila, okuyinto ngokweqile ngenqubo. Lesi sangoma ukuphendula iba ane-alikhali. Okuqukethwe iba umbala owolintshi ekuhlolweni tube.

Izici

Njengoba ksantoproteinovaya ngokuthi ukuphendula qualitative ukuba nitric acid ngenxa yesenzo amaprotheni, isampuli wenziwa ngaphansi kuhlanganisiwe fume kwenzakalani. Ukugcina zonke izinyathelo zokuphepha lapho esebenza kanzulu izinto caustic.

Ukukhululwa okuqukethwe kusukela tube kungenzeka ngesikhathi senqubo Ukushisa okumelwe icatshangelwe lapho kokulilungisa e umnikazi bese ukhetha ukuthambekela.

Ishayela of kanzulu nitric acid soda caustic kufanele kuphela usebenzisa ingilazi pipette futhi amagonandoda iraba pipette kungavunyelwa umlomo.

ukuphendula Isihlobo nge phenol

Ngokucaca, inqubo kanye isiqinisekiso khona amaqembu phenyl Kwenziwa nge efanayo isampula hydroxybenzene.

I tube okwenza 2 ml we phenol buhlanjululwe ke kancane kancane kanye odongeni, ingezwe 2 ml kanzulu nitric acid. Isixazululo yabekwa Ukushisa, lapho kuba ephuzi. Cishe le ndlela yokusabela izinga khona iringi benzene.

Inqubo hydroxybenzene nitrated nge nitric acid kulandele kumiswa nenye p-nitrophenol futhi o-nitrophenol e amaphesenti 15 kuya 35.

Ukuqhathanisa uhlolo gelatin

Ukuze afakazele ukuthi ksantoproteinovaya yokusabela kuya amaprotheni wembula acid acid ne isakhiwo elimnandi, sebenzisa amaprotheni ukuthi akudingeki iqembu phenolic.

Ngo ngamapayipi ehlanzekile senziwa 1% isixazululo gelatin ku-inani 2 ml. Ukuze lokhu wanezela mayelana 9 amathonsi kanzulu nitric acid. Ingxube kwaphumela uyabaselwa. Isixazululo awuzange enombala ophuzi, okuyinto ebonisa ukungabi amino acid nge isakhiwo elimnandi. Ngezinye izikhathi kukhona yellowing kancane imvelo ngenxa yokubakhona wamaprotheni amanyela.

chemical zibalo

Esikhathini ezimbili ukuphendula siphela ksantoproteinovaya amaprotheni. Formula wokuqala esigabeni nitration inqubo uchaza acid acid molecule usebenzisa kanzulu nitric acid.

Isibonelo salokhu okunamathiselwe leqembu nitro le tyrosine ukwakha nitrotyrosine futhi dinitrotirozina. Esimweni sokuqala ukuba iringi benzene unamathele eyodwa CHA ₂ -radical, futhi enkompolo lesibili athomu amabili e-hydrogen esikhundleni CHA _2. Chemical ifomula ksantoproteinovaya tyrosine ukuphendula emelelwa ukuphendula nitric acid ukwakha nitrotyrosine-molecule.

inqubo Nitration kuhambisana ushintsho Umbala kungekho nto enemibala emihle e ithoni ophuzi. Njengoba sifeza lesi ukuphendula namaprotheni aqukethe acid acid izinsalela phenylalanine noma tryptophan, futhi umbala izinguquko isixazululo.

Isinyathelo sesibili siwukuthi umkhiqizo ukusabela nitration ka tyrosine amangqamuzana, ikakhulukazi nitrotyrosine, nge ammonium noma sodium hydroxide. Imiphumela iba yinhlanganisela yokukhandleka sodium noma ammonium usawoti lapho umbala ophuzi. is ukusabela okunjalo ezihlobene kungenzeka nitrotyrosine molecule ukuhambisa ifomu quinoid. Ngemva kwalokho usawoti kwakhiwa therefrom nitronic acid, inohlelo quinone izibopho conjugated double.

Iphela kanjalo iminyaka ksantoproteinovaya yokusabela kuya amaprotheni. I kwesibalo ethulwa ngenhla Isinyathelo sesibili.

Imiphumela

Phakathi ukuhlaziywa eziwuketshezi eziqukethwe amashubhu ezintathu, ikhambi inkomba buhlanjululwe phenol. Izinto nge ring benzene ukunikeza ukuphendula qualitative nge nitric acid. Ngenxa yalokho, lishintsha umbala yesixazululo.

Njengoba yaziwa, gelatin yakhiwa ifomu hydrolyzed collagen. I amaprotheni ayiqukethe isakhiwo elimnandi we amino acid. Alukho ushintsho ngombala medium ukusabela nge acid.

Ngekhulu lesithathu test tube kukhona ukusabela okuhle ngamaprotheni ksantoproteinovaya. Isiphetho kungenziwa shi kanje: zonke amaprotheni ne isakhiwo elimnandi, noma kungaba iqembu phenyl noma i ring indole, banikezwe ushintsho umbala yesixazululo. Lokhu kungenxa kumiswa nitro compounds nge umbala ophuzi.

Kuphethe yokusabela umbala kufakazela khona izakhiwo ezihlukahlukene zamakhemikhali amino acid kanye namaprotheni. ISIBONELO gelatin ebonisa ukuthi ukubunjwa yayo ihlanganisa aminocarboxylic acid ungayembethe iqembu phenyl noma isakhiwo eyindilinga.

Nge ksantoproteinovaya ukuphendula zingachazwa ngezinqubo yellowing kwesikhumba lapho isetshenziswa kube nitric acid eqinile. Nombala ofanayo iyoba ubisi frothy phakathi ukuhlaziya yakhe efanayo.

Ngo emitholampilo laboratory umkhuba lokhu isampula umbala alisetshenziswa ngenxa ukuthola amaprotheni ku umchamo. Lokhu kungenxa umbala ophuzi we umchamo uqobo.

Ksantoproteinovaya ukuphendula babehlukaniswa kakhulu esetshenziselwa quantification amino acid ezifana tryptophan futhi tyrosine, njengengxenye amaprotheni ahlukahlukene.