Li-peptide ke sehlopha sa metsoako e entsoeng ka ho hokahana ha li-amino acid tse ngata ka li-bond tsa peptide.Li fumaneha hohle linthong tse phelang.Ho fihlela joale, li-peptide tse mashome a likete li fumanoe linthong tse phelang.Li-peptide li bapala karolo ea bohlokoa ho laola mesebetsi ea ts'ebetso ea litsamaiso tse fapaneng, litho, lisele le lisele le mesebetsing ea bophelo, 'me hangata li sebelisoa tlhahlobong ea ts'ebetso, lipatlisiso tsa antibody, nts'etsopele ea lithethefatsi le likarolo tse ling.Ka nts'etsopele ea theknoloji ea biotechnology le peptide synthesis, meriana ea peptide e ntse e eketseha le ho sebelisoa tleliniking.
Ho na le mefuta e mengata e fapaneng ea liphetoho tsa peptide, tse ka aroloang habonolo feela ka phetoho ea poso le phetoho ea ts'ebetso (ho sebelisa phetoho e nkiloeng ea amino acid), le phetoho ea N-terminal, phetoho ea C-terminal, phetoho ea ketane ea mahlakoreng, phetoho ea amino acid, phetoho ea skeleton, joalo-joalo, ho itšetlehile ka sebaka sa phetoho (setšoantšo sa 1).E le mokhoa oa bohlokoa oa ho fetola sebopeho sa ketane kapa lihlopha tsa ketane ea mahlakoreng a liketane tsa peptide, phetoho ea peptide e ka fetola ka nepo thepa ea 'mele le ea lik'hemik'hale ea metsoako ea peptide, ea eketsa ho qhibiliha ha metsi, ho lelefatsa nako ea ts'ebetso ka vivo, ho fetola phepelo ea bona ea tlhaho, ho felisa immunogenicity. , fokotsa litla-morao tse chefo, joalo-joalo pampiring ena, ho hlahisoa maano a 'maloa a maholo a ho fetola peptide le litšobotsi tsa ona.
1. Ho tsamaea ka baesekele
Li-peptide tsa cyclic li na le lits'ebetso tse ngata ho biomedicine, 'me li-peptide tse ngata tsa tlhaho tse nang le ts'ebetso ea bioloji ke li-peptide tsa cyclic.Hobane li-peptide tsa cyclic li na le ho thatafala ho feta li-peptide tse melang, li hanana haholo le tsamaiso ea tšilo ea lijo, li khona ho phela ka har'a tšilo ea lijo, 'me li bonts'a kamano e matla bakeng sa li-receptor tse shebiloeng.Cyclization ke mokhoa o tobileng ka ho fetesisa oa ho kopanya li-peptide tsa cyclic, haholo-holo bakeng sa li-peptide tse nang le skeleton e kholo ea sebopeho.Ho ea ka mokhoa oa cyclization, e ka aroloa ka mofuta oa ketane ea lehlakore-lehlakoreng, mofuta oa ketane ea lehlakore, mofuta oa terminal - mofuta oa ho qetela (mofuta oa ho qetela).
(1) sidechain-to-sidechain
Mofuta o atileng haholo oa ketane ea mahlakoreng ho ea ho ketane ea lehlakore ke borokho ba disulfide lipakeng tsa masala a cysteine.cyclization ena e hlahisoa ke para ea masala a cysteine a sirelelitsoeng ebe o oxidized ho theha maqhama a disulfide.Polycyclic synthesis e ka finyelloa ka ho tlosoa ho khethehileng ha lihlopha tsa ts'ireletso ea sulfhydryl.Cyclization e ka etsoa ka motsoako oa post-dissociation kapa ka resin ea pre-dissociation.Ho potoloha ka li-resin ho ka 'na ha se ke ha sebetsa hantle ho feta solvent cyclization hobane li-peptide tse holim'a li-resin ha li khone ho etsa li-conformations tsa cyclified.Mofuta o mong oa ketane ea mahlakoreng - cyclization ea ketane ea mahlakoreng ke sebopeho sa sebopeho sa amide lipakeng tsa asiti ea aspartic kapa masala a asiti ea glutamic le amino acid, e hlokang hore sehlopha sa ts'ireletso ea ketane ea mahlakoreng se khone ho tlosoa ka mokhoa o ikhethileng ho polypeptide. holim'a resin kapa ka mor'a ho arohana.Mofuta oa boraro oa ketane ea mahlakoreng - cyclization ea mahlakoreng ke ho thehoa ha diphenyl ethers ka tyrosine kapa p-hydroxyphenylglycine.Mofuta ona oa ho palama libaesekele lihlahisoa tsa tlhaho o fumanoa feela lihlahisoa tse nyane, mme lihlahisoa tsa cyclization hangata li na le boleng ba bongaka bo ka bang teng.Ho lokisoa ha metsoako ena ho hloka maemo a ikhethang a karabelo, kahoo ha a sebelisoe hangata ha ho etsoa li-peptide tse tloaelehileng.
(2) terminal-to-sidechain
Theminale-side chain cyclization hangata e kenyelletsa C-terminal le sehlopha sa amino sa ketane ea lysine kapa ornithine, kapa N-terminal e nang le aspartic acid kapa glutamic acid side chain.Polypeptide cyclization e 'ngoe e etsoa ka ho theha maqhama a ether lipakeng tsa terminal C le liketane tsa lehlakore tsa serine kapa tsa threonine.
(3) Mofuta oa terminal kapa hlooho-to-mohatla
Li-polypeptides tsa Chain li ka tsamaisoa ka baesekele ka har'a sehlapi kapa tsa ts'oaroa holim'a resin ka lehlakoreng la ketane ea cyclation.Maemo a tlase a li-peptide a lokela ho sebelisoa ho solvent centralization ho qoba oligomerization ea li-peptide.Sehlahisoa sa polypeptide e entsoeng ka hlooho-to-mohatla e itšetlehile ka tatellano ea polypeptide ea ketane.Ka hona, pele o lokisa li-peptide tsa cyclic ka tekanyo e kholo, laeborari ea li-peptide tse ka khonehang tse kentsoeng li lokela ho qala pele ho theoa, e lateloe ke cyclization ho fumana tatellano e nang le litholoana tse ntle.
2. N-methylation
N-methylation qalong e hlaha ho li-peptide tsa tlhaho 'me e kenngoa ka har'a peptide synthesis ho thibela ho thehoa ha litlamo tsa hydrogen, ka tsela eo li etsa hore li-peptide li se ke tsa hlola li e-ba le likokoana-hloko le ho tlosoa.Motsoako oa li-peptide tse sebelisang lihlahisoa tsa N-methylated amino acid ke mokhoa oa bohlokoahali.Ho feta moo, karabelo ea Mitsunobu ea N-(2-nitrobenzene sulfonyl chloride) polypeptide-resin intermediates le methanol le eona e ka sebelisoa.Mokhoa ona o sebelisitsoe ho lokisa lilaebrari tsa cyclic peptide tse nang le N-methylated amino acid.
3. Phosphorylation
Phosphorylation ke e 'ngoe ea liphetolelo tse atileng haholo tsa morao-rao tsa tlhaho.Liseleng tsa motho, ho feta 30% ea liprotheine ke phosphorylated.Phosphorylation, haholo-holo phosphorylation e fetolehang, e phetha karolo ea bohlokoa ho laola mekhoa e mengata ea lisele, joalo ka phetisetso ea matšoao, polelo ea liphatsa tsa lefutso, potoloho ea lisele le taolo ea cytoskeleton, le apoptosis.
Phosphorylation e ka bonoa mesaletsa ea amino acid e fapaneng, empa sepheo se tloaelehileng sa phosphorylation ke masala a serine, threonine le tyrosine.Phosphotyrosine, phosphothreonine, le phosphoserine derivatives e ka kenngoa ka har'a li-peptide nakong ea ho kopanya kapa ea thehoa ka mor'a peptide synthesis.Phosphorylation e khethiloeng e ka finyelloa ho sebelisoa masala a serine, threonine, le tyrosine tse tlosang lihlopha tse sireletsang ka boikhethelo.Li-reagents tse ling tsa phosphorylation le tsona li ka hlahisa lihlopha tsa asiti ea phosphoric ka har'a polypeptide ka phetoho ea poso.Lilemong tsa morao tjena, phosphorylation e khethehileng ea sebaka sa lysine e 'nile ea finyelloa ka ho sebelisa mokhoa o khethollang lik'hemik'hale oa Staudinger-phosphite (Setšoantšo sa 3).
4. Myristoylation le palmitoylation
Acylation ea N-terminal e nang le mafura a acid e lumella li-peptide kapa liprotheine ho tlama lera la sele.Tatellano ea myridamoylated ho N-terminal e thusa Src family protein kinases le reverse transcriptase Gaq proteins hore e lebisoe ho tlama liseleng.Myristic acid e ne e amahanngoa le N-terminal ea resin-polypeptide e sebelisa mekhoa e tloaelehileng ea ho kopanya, 'me sephetho sa lipopeptide se ka aroloa tlas'a maemo a tloaelehileng' me sa hloekisoa ke RP-HPLC.
5. Glycosylation
Li-Glycopeptides tse kang vancomycin le teicolanin ke lithibela-mafu tsa bohlokoa bakeng sa ho phekola mafu a baktheria a hananang le lithethefatsi, ’me hangata li-glycopeptide tse ling li sebelisoa ho susumetsa sesole sa ’mele.Ho feta moo, kaha li-antigen tse ngata tsa likokoana-hloko li na le glycosylated, ho bohlokoa haholo ho ithuta ka glycopeptides bakeng sa ho ntlafatsa phello ea phekolo ea tšoaetso.Ka lehlakoreng le leng, ho fumanoe hore liprotheine tse ka lera la lisele tsa tumor li bonts'a glycosylation e sa tloaelehang, e etsang hore glycopeptides e be le karolo ea bohlokoa ho mofets'e le lipatlisisong tsa ts'ireletso ea 'mele oa ho itšireletsa mafung.Li-Glycopeptides li lokisoa ka mokhoa oa Fmoc/t-Bu.Masala a glycosylated, joalo ka threonine le serine, hangata a kenngoa ka har'a polypeptides ke pentafluorophenol ester activated fMOCs ho sireletsa glycosylated amino acid.
6. Isoprene
Isopentadienylation e etsahala ka masala a cysteine ka har'a ketane e ka thōko haufi le C-terminal.Protein isoprene e ka ntlafatsa kamano ea lera la sele le ho theha tšebelisano ea protheine le protheine.Liprotheine tse nang le Isopentadienate li kenyelletsa tyrosine phosphatase, GTase e nyane, limolek'hule tsa cochaperone, lamina ea nyutlelie, le liprotheine tse tlamang tsa centromeric.Isoprene polypeptides e ka lokisoa ho sebelisoa isoprene holim'a li-resin kapa ka ho hlahisa lihlahisoa tsa cysteine.
7. Phetoho ea polyethylene glycol (PEG).
Phetoho ea PEG e ka sebelisoa ho ntlafatsa botsitso ba protheine hydrolytic, biodistribution le solubility ea peptide.Ho hlahisoa ha liketane tsa PEG ho li-peptide ho ka ntlafatsa thepa ea bona ea pharmacological hape ho thibela hydrolysis ea li-peptide ka li-enzyme tsa proteolytic.Li-peptide tsa PEG li feta karolong ea glomerular capillary cross habonolo ho feta li-peptide tse tloaelehileng, 'me li fokotsa haholo ho hloekisoa ha renal.Ka lebaka la nako e telele ea halofo ea bophelo ea li-peptide tsa PEG ka har'a vivo, boemo bo tloaelehileng ba kalafo bo ka bolokoa ka litekanyetso tse tlase le lithethefatsi tsa peptide tse fokolang khafetsa.Leha ho le joalo, phetoho ea PEG le eona e na le litlamorao tse mpe.Palo e kholo ea PEG e thibela enzyme ho theola peptide hape e fokotsa ho tlama ha peptide ho se amohelang sepheo.Empa kamano e tlase ea PEG peptides hangata e koaheloa ke halofo ea bophelo ba bona bo bolelele ba pharmacokinetic, mme ka ho ba teng 'meleng nako e telele, li-peptide tsa PEG li na le monyetla o moholo oa ho kenngoa liseleng tse shebiloeng.Ka hona, litlhaloso tsa PEG polymer li lokela ho ntlafatsoa bakeng sa liphetho tse nepahetseng.Ka lehlakoreng le leng, li-peptide tsa PEG li bokellana sebeteng ka lebaka la ho fokotseha ha renal clearance, ho fella ka macromolecular syndrome.Ka hona, liphetoho tsa PEG li hloka ho raloa ka hloko haholoanyane ha li-peptide li sebelisoa bakeng sa tlhahlobo ea lithethefatsi.
Lihlopha tse tloaelehileng tsa liphetoho tsa PEG modifiers li ka akaretsa ka mokhoa o latelang: 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 (e kang propional-ald, butyrALD), acrylic base (-acrylate-acrl), azido-azide, biotinyl - Biotin, Fluorescein, glutaryl -GA, Acrylate Hydrazide, alkyne-alkyne, p-toluenesulfonate -OTs, succinimide succinate -SS, joalo-joalo Lihlahisoa tsa PEG tse nang le carboxylic acid li ka kopanngoa le li-amine tsa n-terminal kapa liketane tsa mahlakoreng a lysine.Amino-activated PEG e ka kopanngoa le liketane tsa mahlakoreng tsa asiti ea aspartic kapa glutamic acid.PEG e kentsoeng hampe e ka kopanngoa le mercaptan ea liketane tse ka thōko tsa cysteine tse sirelelitsoeng ka ho feletseng [11].Liphetoho tsa PEG hangata li hlophisoa ka tsela e latelang (hlokomela: mPEG ke methoxy-PEG, CH3O-(CH2CH2O)n-CH2CH2-OH) :
(1) ketane e otlolohileng ea PEG modifier
mPEG-SC, mPEG-SCM, mPEG-SPA, mPEG-OTs, mPEG-SH, mPEG-ALD, mPEG-butyrALD, mPEG-SS
(2) phetoho ea PEG e sa sebetseng habeli
HCOO-PEG-COOH, NH2-PEG-NH2, OH-PEG-COOH, OH-PEG-NH2, HCl·NH2-PEG-COOH, MAL-PEG-NHS
(3) branching PEG modifier
(mPEG)2-NHS, (mPEG)2-ALD, (mPEG)2-NH2, (mPEG)2-MAL
8. Biotinization
Biotin e ka tlamahanngoa ka matla le avidin kapa streptavidin, 'me matla a tlamang a bile a le haufi le tlamo ea covalent.Li-peptide tse ngotsoeng ka Biotin li sebelisoa hangata ho immunoassay, histocytochemistry, le fluorescence-based flow cytometry.Li-antibiotin tse ngolisitsoeng li ka boela tsa sebelisoa ho tlama li-peptide tsa biotinylated.Li-labels tsa Biotin hangata li khomaretsoe ho ketane ea lehlakoreng la lysine kapa N terminal.6-aminocaproic acid hangata e sebelisoa e le tlamo lipakeng tsa peptide le biotin.Bond e fetoha habonolo ha e tlama ho substrate mme e tlama hantle ka ho ba teng ha tšitiso ea steric.
9. Ho ngola ka fluorescent
Ho ngola ka fluorescent ho ka sebelisoa ho latela polypeptide liseleng tse phelang le ho ithuta li-enzyme le mekhoa ea ts'ebetso.Tryptophan (Trp) ke fluorescent, kahoo e ka sebelisoa bakeng sa ho ngola ka hare ho naha.Sekhahla sa tlhahiso ea tryptophan se ipapisitse le tikoloho ea peripheral mme se fokotseha ka ho fokotseha ha polarity ea solvent, thepa e thusang ho bona sebopeho sa peptide le ho tlama li-receptor.Tryptophan fluorescence e ka tima ka protonated aspartic acid le glutamic acid, e ka fokotsang tšebeliso ea eona.Sehlopha sa Dansyl chloride (Dansyl) se na le fluorescent haholo ha se tlameletsoe sehlopheng sa amino 'me hangata se sebelisoa e le leibole ea fluorescent bakeng sa amino acid kapa liprotheine.
Fluorescence resonance Energy conversion (FRET) e thusa lithutong tsa enzyme.Ha FRET e sebelisoa, substrate polypeptide hangata e na le sehlopha sa fluorescence-labeling le sehlopha sa fluorescence-quenching.Lihlopha tse ngotsoeng ka fluorescent li tingoa ke quencher ka ho fetisa matla a se nang photon.Ha peptide e aroloa ho enzyme eo ho buuoang ka eona, sehlopha se ngotseng se ntša fluorescence.
10. Cage polypeptides
Li-cage peptide li na le lihlopha tse sirelletsang tse ka tlosoang tse sirelletsang peptide hore e se ke ea tlama ho receptor.Ha peptide e pepesehetse mahlaseli a kotsi a UV, e ea sebetsa, 'me e khutlisetsa kamano ea eona ho receptor.Hobane ts'ebetso ena ea optical e ka laoloa ho latela nako, amplitude, kapa sebaka, li-cage peptide li ka sebelisoa ho ithuta karabelo e etsahalang liseleng.Lihlopha tsa ts'ireletso tse sebelisoang haholo bakeng sa cage polypeptides ke lihlopha tsa 2-nitrobenzyl le lihlahisoa tsa tsona, tse ka hlahisoang ka peptide synthesis ka li-derivatives tse sireletsang tsa amino acid.Lintho tse tsoang ho amino acid tse entsoeng ke lysine, cysteine, serine, le tyrosine.Li-derivatives tsa aspartate le glutamate, leha ho le joalo, ha li sebelisoe hangata ka lebaka la ts'oaetso ea tsona ea ho palama libaesekele nakong ea peptide synthesis le dissociation.
11. Polyantigenic peptide (MAP)
Li-peptide tse khutšoane hangata ha li sireletsehe 'me li tlameha ho kopanngoa le liprotheine tse tsamaisang ho hlahisa li-antibodies.Polyantigenic peptide (MAP) e entsoe ka li-peptide tse ngata tse ts'oanang tse hokahaneng le lysine nuclei, tse ka hlalosang ka ho hlaka li-immunogens tse phahameng 'me li ka sebelisoa ho lokisa li-couplets tsa protheine e tsamaisang peptide.Li-polypeptides tsa MAP li ka kopanngoa ke motsoako o tiileng oa karolo ho resin ea MAP.Leha ho le joalo, khokahanyo e sa fellang e fella ka liketane tsa peptide tse sieo kapa tse fokolitsoeng makaleng a mang, ka hona ha e bontše thepa ea MAP polypeptide ea mantlha.Joalo ka mokhoa o mong, li-peptide li ka lokisoa le ho hloekisoa ka thoko ebe li kopanngoa le MAP.Tatelano ea peptide e hoketsoeng mokokotlong oa peptide e hlalosoa hantle ebile e tsebahala habonolo ka boima ba pono.
Qetello
Phetoho ea peptide ke mokhoa oa bohlokoa oa ho qapa li-peptide.Li-peptide tse fetotsoeng ka lik'hemik'hale ha li khone feela ho boloka ts'ebetso e phahameng ea likokoana-hloko, empa hape li qoba ka katleho litšitiso tsa immunogenicity le chefo.Ka nako e ts'oanang, phetoho ea lik'hemik'hale e ka etsa hore li-peptide li be le thepa e ncha e ntle haholo.Lilemong tsa morao tjena, mokhoa oa ts'ebetso ea CH bakeng sa phetoho ea morao-rao ea polypeptide e 'nile ea ntlafatsoa ka potlako,' me liphello tse ngata tsa bohlokoa li finyelitsoe.
Nako ea poso: Mar-20-2023