Peptides rukuni ne na mahadi da aka kafa ta hanyar haɗin amino acid da yawa ta hanyar haɗin peptide.Suna da yawa a cikin halittu masu rai.Ya zuwa yanzu, an samu dubunnan peptides a cikin halittu masu rai.Peptides suna taka muhimmiyar rawa wajen daidaita ayyukan ayyuka na tsarin daban-daban, gabobin jiki, kyallen takarda da sel da kuma cikin ayyukan rayuwa, kuma galibi ana amfani da su a cikin bincike na aiki, bincike na antibody, ci gaban ƙwayoyi da sauran fannoni.Tare da haɓaka fasahar kere-kere da fasahar haɗin peptide, an haɓaka ƙarin magungunan peptide kuma ana amfani da su a asibiti.
Akwai nau'ikan gyare-gyare na peptide iri-iri, waɗanda kawai za'a iya raba su zuwa gyare-gyare na baya da gyare-gyaren tsari (ta amfani da gyare-gyaren amino acid da aka samo), da gyaran N-terminal, gyare-gyaren C-terminal, gyaran sarkar gefe, gyaran amino acid, gyaran kwarangwal. da sauransu, dangane da wurin gyarawa (Hoto 1).A matsayin hanya mai mahimmanci don canza babban tsarin sarkar ko ƙungiyoyin sassan sassan peptide, gyare-gyaren peptide zai iya canza yanayin jiki da sinadarai na mahadi na peptide yadda ya kamata, ƙara ruwa mai narkewa, tsawaita lokacin aiki a cikin vivo, canza rarraba kwayoyin su, kawar da immunogenicity. , rage illa masu guba, da dai sauransu. A cikin wannan takarda, an gabatar da manyan dabarun gyara peptide da dama da halayen su.
1. Yin hawan keke
Cyclic peptides suna da aikace-aikace da yawa a cikin biomedicine, kuma yawancin peptides na halitta tare da ayyukan nazarin halittu sune peptides na cyclic.Saboda peptides na cyclic sun kasance sun fi tsayi fiye da peptides na layi, suna da matukar juriya ga tsarin narkewa, suna iya rayuwa a cikin tsarin narkewa, kuma suna nuna dangantaka mai karfi ga masu karɓa.Cyclization ita ce hanya mafi kai tsaye don haɗa peptides na cyclic, musamman ga peptides tare da manyan kwarangwal na tsari.Dangane da yanayin hawan keke, ana iya raba shi zuwa nau'in sarkar gefe-gefe, tasha - nau'in sarkar gefe, tasha - nau'in tasha (nau'in ƙarshen zuwa ƙarshen).
(1) sikelin-zuwa-gefe
Mafi yawan nau'in sarkar gefe zuwa hawan keke na gefe shine haɗin disulfide tsakanin ragowar cysteine.Ana gabatar da wannan hawan keke ta hanyar wasu ɓangarori biyu na cysteine da ake kare su sannan kuma an sanya su da iskar oxygen don samar da haɗin gwiwar disulfide.Ana iya samun haɗin polycyclic ta zaɓin cire ƙungiyoyin kariya na sulfhydryl.Ana iya yin keken keke ko dai a cikin wani kaushi bayan rabuwa ko kuma a kan resin da aka riga aka raba.Cyclization a kan resins na iya zama ƙasa da tasiri fiye da cyclization na ƙarfi saboda peptides akan resins ba sa samar da cyclified conformations.Wani nau'in sarkar gefe - hawan sarkar gefe shine samuwar tsari na amide tsakanin ragowar aspartic acid ko glutamic acid da amino acid na tushe, wanda ke buƙatar rukunin kariya daga sarkar gefe dole ne a iya zaɓin cire shi daga polypeptide ko dai. a kan guduro ko bayan rabuwa.Nau'in nau'i na uku na gefen-sarkar - cyclization sarkar gefe shine samuwar diphenyl ethers ta tyrosine ko p-hydroxyphenylglycine.Wannan nau'in hawan keke a cikin samfuran halitta ana samunsa ne kawai a cikin samfuran ƙwayoyin cuta, kuma samfuran hawan keke galibi suna da yuwuwar ƙimar magani.Shirye-shiryen waɗannan mahadi yana buƙatar yanayi na musamman, don haka ba a amfani da su sau da yawa a cikin kira na peptides na al'ada.
(2) Terminal-to-sidechain
Kewayon sarkar gefe-gefe yawanci ya ƙunshi C-terminal tare da rukunin amino na lysine ko sarkar gefen ornithine, ko N-terminal tare da sarkar gefen aspartic acid ko glutamic acid.Ana yin wasu cyclization na polypeptide ta hanyar samar da ether bond tsakanin m C da serine ko threonine gefen sarƙoƙi.
(3) Terminal ko nau'in kai-zuwa wutsiya
Za'a iya yin hawan sarkar polypeptides ko dai a cikin keken keke a cikin wani ƙarfi ko daidaitawa akan guduro ta hanyar hawan sarkar gefe.Ya kamata a yi amfani da ƙananan ƙwayoyin peptides a cikin tsaka-tsakin ƙarfi don guje wa oligomerization na peptides.Samuwar zoben roba polypeptide kai-da- wutsiya ya dogara da jerin sarkar polypeptide.Sabili da haka, kafin shirya peptides na cyclic a kan babban sikelin, ya kamata a fara ƙirƙirar ɗakin karatu na yuwuwar peptides na gubar da aka ɗaure, sannan a bi da keke don nemo jerin tare da sakamako mafi kyau.
2. N-methylation
N-methylation da farko yana faruwa a cikin peptides na halitta kuma an gabatar da shi a cikin haɗin peptide don hana samuwar haɗin hydrogen, ta haka yana sa peptides ya fi tsayayya ga biodegradation da sharewa.Haɓaka peptides ta amfani da abubuwan amino acid N-methylated shine hanya mafi mahimmanci.Bugu da ƙari, Mitsunobu dauki na N- (2-nitrobenzene sulfonyl chloride) polypeptide-resin intermediates tare da methanol kuma za a iya amfani da.An yi amfani da wannan hanyar don shirya ɗakunan karatu na peptide na cyclic masu ɗauke da amino acid N-methylated.
3. Phosphorylation
Phosphorylation yana ɗaya daga cikin gyare-gyaren da aka fi sani bayan fassarar a yanayi.A cikin kwayoyin jikin mutum, fiye da 30% na sunadaran sunadaran phosphorylated.Phosphorylation, musamman reversible phosphorylation, taka muhimmiyar rawa wajen sarrafa da yawa salon salula tafiyar matakai, kamar sigina sigina, gene expression, cell sake zagayowar da cytoskeleton tsari, da apoptosis.
Ana iya lura da phosphorylation a ragowar amino acid iri-iri, amma mafi yawan maƙasudin phosphorylation sune ragowar serine, threonine, da tyrosine.Phosphotyrosine, phosphothreonine, da abubuwan phosphoserine za a iya shigar da su a cikin peptides yayin da ake kira ko kuma an kafa su bayan haɗin peptide.Za'a iya samun zaɓin phosphorylation ta amfani da ragowar serine, threonine, da tyrosine waɗanda ke zaɓin cire ƙungiyoyin kariya.Wasu reagents na phosphorylation kuma na iya gabatar da ƙungiyoyin phosphoric acid a cikin polypeptide ta bayan gyara.A cikin 'yan shekarun nan, an sami phosphorylation na musamman na lysine ta hanyar amfani da sinadarai na Staudinger-phosphite dauki (Hoto 3).
4. Myristoylation da palmitoylation
Acylation na N-terminal tare da fatty acid yana ba da damar peptides ko sunadaran don ɗaure ga membranes tantanin halitta.Jerin myridamoylated akan N-terminal yana bawa Src protein kinases na iyali da juyar da sunadaran Gaq da za a yi niyya don ɗaure ga membranes cell.An haɗa Myristic acid zuwa N-terminal na resin-polypeptide ta yin amfani da daidaitattun halayen haɗin gwiwa, kuma sakamakon lipopeptide zai iya rabuwa a ƙarƙashin daidaitattun yanayi kuma an tsarkake shi ta RP-HPLC.
5. Glycosylation
Glycopeptides irin su vancomycin da teicolanin sune mahimman maganin rigakafi don maganin cututtukan ƙwayoyin cuta masu jure wa ƙwayoyi, kuma ana amfani da sauran glycopeptides don tada tsarin rigakafi.Bugu da ƙari, tun da yawancin antigens na microbial suna glycosylated, yana da mahimmanci don nazarin glycopeptides don inganta tasirin warkewa na kamuwa da cuta.A gefe guda kuma, an gano cewa sunadaran da ke jikin membrane cell na ƙwayoyin tumor suna nuna glycosylation mara kyau, wanda ya sa glycopeptides ke taka muhimmiyar rawa wajen bincike na kare kariya daga ciwon daji.Ana shirya Glycopeptides ta hanyar Fmoc/t-Bu.Ragowar Glycosylated, kamar threonine da serine, galibi ana shigar dasu cikin polypeptides ta hanyar pentafluorophenol ester kunna fMOCs don kare amino acid glycosylated.
6. Isoprene
Isopentadienylation yana faruwa akan ragowar cysteine a cikin sassan gefe kusa da C-terminal.Protein isoprene na iya inganta alaƙar membrane na sel kuma ya samar da hulɗar furotin-gina jiki.Sunadaran da aka ware sun haɗa da tyrosine phosphatase, ƙananan GTase, kwayoyin cochaperone, lamina na nukiliya, da kuma sunadaran dauri na centromeric.Ana iya shirya polypeptides na isoprene ta amfani da isoprene akan resins ko ta hanyar gabatar da abubuwan cysteine.
7. Polyethylene glycol (PEG) gyara
Ana iya amfani da gyare-gyaren PEG don inganta zaman lafiyar furotin hydrolytic, rarraba biodistribution da peptide solubility.Gabatar da sarƙoƙin PEG zuwa peptides na iya haɓaka kaddarorin su na harhada magunguna da kuma hana hydrolysis na peptides ta hanyar enzymes proteolytic.PEG peptides suna wucewa ta sashin glomerular capillary giciye cikin sauƙi fiye da peptides na yau da kullun, suna raguwa sosai.Saboda tsawaita rabin rayuwar PEG peptides a cikin vivo, ana iya kiyaye matakin jiyya na yau da kullun tare da ƙananan allurai da ƙarancin magungunan peptide.Koyaya, gyaran PEG shima yana da mummunan tasiri.Yawancin PEG yana hana enzyme daga lalata peptide kuma yana rage daurin peptide ga mai karɓar manufa.Amma ƙarancin kusancin PEG peptides yawanci ana kashe su ta tsawon rabin rayuwar su na pharmacokinetic, kuma ta kasancewa a cikin jiki tsawon lokaci, peptides na PEG suna da yuwuwar shiga cikin kyallen takarda.Don haka, ya kamata a inganta ƙayyadaddun bayanai na PEG polymer don ingantacciyar sakamako.A gefe guda kuma, peptides na PEG suna taruwa a cikin hanta saboda raguwar raguwar koda, yana haifar da ciwo na macromolecular.Don haka, ana buƙatar gyare-gyaren PEG a hankali sosai lokacin da ake amfani da peptides don gwajin ƙwayoyi.
Ƙungiyoyin gyare-gyare na yau da kullum na masu gyara PEG za a iya taƙaita su kamar haka: Amino (-amine) -NH2, aminomethyl-Ch2-NH2, hydroxy-OH, carboxy-Cooh, sulfhydryl (-Thiol) -SH, Maleimide -MAL, succinimide carbonate - SC, succinimide acetate -SCM, succinimide propionate -SPA, n-hydroxysuccinimide -NHS, Acrylate-ch2ch2cooh, aldehyde -CHO (irin su propional-ald, butyrALD), acrylic tushe (-acrylate-acrl), azido-azide, biotinyl - Biotin, Fluorescein, glutaryl -GA, Acrylate Hydrazide, alkyne-alkyne, p-toluenesulfonate -OTs, succinimide succinate -SS, da sauransu. Abubuwan PEG tare da acid carboxylic ana iya haɗa su zuwa amines n-terminal ko sarƙoƙin gefen lysine.Amino-activated PEG ana iya haɗa shi zuwa aspartic acid ko sarƙoƙin gefen glutamic acid.PEG mai kunnawa mara kyau ana iya haɗa shi zuwa mercaptan na sarƙoƙin gefen cysteine mara kyau [11].Ana rarraba masu gyara PEG kamar haka (bayanin kula: mPEG shine methoxy-PEG, CH3O-(CH2CH2O) n-CH2CH2-OH):
(1) madaidaiciyar sarkar PEG mai gyara
mPEG-SC, mPEG-SCM, mPEG-SPA, mPEG-OTs, mPEG-SH, mPEG-ALD, mPEG-butyrALD, mPEG-SS
(2) Mai gyara PEG mai aiki biyu
HCOO-PEG-COOH, NH2-PEG-NH2, OH-PEG-COOH, OH-PEG-NH2, HCl·NH2-PEG-COOH, MAL-PEG-NHS
(3) Mai gyara PEG mai reshe
(mPEG) 2-NHS, (mPEG) 2-ALD, (mPEG) 2-NH2, (mPEG) 2-MAL
8. Biotinization
Ana iya ɗaure Biotin da ƙarfi tare da avidin ko streptavidin, kuma ƙarfin ɗaurin yana kusa da haɗin gwiwa.Ana amfani da peptides masu lakabin biotin a cikin immunoassay, histocytochemistry, da cytometry na kwararar haske.Hakanan ana iya amfani da ƙwayoyin rigakafin ƙwayoyin cuta masu lakabi don ɗaure peptides na biotinylated.Alamun biotin galibi ana haɗe su zuwa sarkar gefen lysine ko tashar N.6-aminocaproic acid ana yawan amfani dashi azaman haɗin gwiwa tsakanin peptides da biotin.Haɗin yana da sassauƙa wajen ɗaure ga ma'auni kuma yana ɗaure mafi kyau a gaban shinge mai tsauri.
9. Alamar walƙiya
Za a iya amfani da lakabin fluorescent don gano polypeptides a cikin sel masu rai da kuma nazarin enzymes da hanyoyin aiki.Tryptophan (Trp) mai kyalli ne, don haka ana iya amfani da shi don yin lakabin ciki.Fitar da bakan tryptophan ya dogara ne akan mahalli na gefe kuma yana raguwa tare da raguwar polarity mai ƙarfi, kayan da ke da amfani don gano tsarin peptide da ɗaure mai karɓa.Tryptophan fluorescence za a iya kashe ta protonated aspartic acid da glutamic acid, wanda zai iya iyakance amfani da shi.Ƙungiyar Dansyl chloride (Dansyl) tana da haske sosai idan an ɗaure ta da ƙungiyar amino kuma galibi ana amfani da ita azaman alamar kyalli don amino acid ko furotin.
Fluorescence resonance Energy canzawa (FRET) yana da amfani don nazarin enzyme.Lokacin da aka yi amfani da FRET, polypeptide substrate yawanci yana ƙunshe da rukuni mai alamar haske da ƙungiyar kashe haske.Ƙungiyoyin da aka yi wa laƙabi suna kashe su ta hanyar mai kashe wuta ta hanyar canjin makamashi mara hoto.Lokacin da peptide ya rabu da enzyme da ake tambaya, rukunin lakabi yana fitar da haske.
10. Cage polypeptides
Cage peptides suna da ƙungiyoyin kariya masu iya cirewa waɗanda ke kare peptide daga ɗaure ga mai karɓa.Lokacin da aka fallasa shi zuwa hasken UV, peptide yana kunna, yana maido da kusancinsa ga mai karɓa.Saboda ana iya sarrafa wannan kunnawar gani gwargwadon lokaci, girman ko wuri, ana iya amfani da peptides keji don nazarin halayen da ke faruwa a cikin sel.Ƙungiyoyin kariya da aka fi amfani da su don polypeptides keji sune ƙungiyoyin 2-nitrobenzyl da abubuwan da suka samo asali, waɗanda za a iya gabatar da su a cikin haɗin peptide ta hanyar abubuwan amino acid masu kariya.Abubuwan da aka samo asali na amino acid sune lysine, cysteine, serine, da tyrosine.Abubuwan da aka samo asali na aspartate da glutamate, duk da haka, ba a saba amfani da su ba saboda lalurarsu ga hawan keke yayin haɗuwar peptide da rarrabawa.
11. Polyantigenic peptide (MAP)
Short peptides yawanci ba su da rigakafi kuma dole ne a haɗa su da sunadarai masu ɗaukar hoto don samar da ƙwayoyin rigakafi.Polyantigenic peptide (MAP) yana kunshe da nau'ikan peptides iri ɗaya da ke da alaƙa da lysine nuclei, waɗanda ke iya bayyana babban ƙarfin immunogens musamman kuma ana iya amfani da su don shirya ma'auratan furotin mai ɗaukar peptide.MAP polypeptides za a iya haɗe ta ta ingantaccen tsarin lokaci akan guduro MAP.Duk da haka, rashin cikar haɗin kai yana haifar da ɓacewa ko yanke sarƙoƙin peptide a wasu rassan kuma don haka baya nuna kaddarorin ainihin MAP polypeptide.A madadin, ana iya shirya peptides kuma a tsarkake su daban sannan a haɗa su zuwa MAP.Matsakaicin peptide da aka haɗe zuwa ainihin peptide yana da ma'ana da kyau kuma cikin sauƙi ana iya siffanta shi ta hanyar spectrometry na taro.
Kammalawa
Gyaran peptide hanya ce mai mahimmanci na zayyana peptides.Abubuwan peptides da aka gyara ta hanyar sinadarai ba za su iya kiyaye manyan ayyukan ilimin halitta kawai ba, har ma yadda ya kamata su guje wa abubuwan da ke tattare da rigakafi da guba.A lokaci guda, gyare-gyaren sinadarai na iya ba da peptides tare da wasu sababbin kyawawan kaddarorin.A cikin 'yan shekarun nan, hanyar CH kunnawa don gyare-gyare na polypeptides an haɓaka da sauri, kuma an sami sakamako mai mahimmanci.
Lokacin aikawa: Maris 20-2023