Peptides yog ib chav kawm ntawm cov tebchaw tsim los ntawm kev sib txuas ntawm ntau cov amino acids los ntawm peptide bonds.Lawv yog ubiquitous nyob rau hauv cov kab mob.Txog tam sim no, kaum tawm txhiab tus peptides tau pom nyob hauv cov kab mob muaj sia.Peptides ua lub luag haujlwm tseem ceeb hauv kev tswj hwm kev ua haujlwm ntawm ntau lub cev, lub cev, cov ntaub so ntswg thiab cov hlwb thiab hauv kev ua haujlwm hauv lub neej, thiab feem ntau siv rau hauv kev tshuaj xyuas kev ua haujlwm, kev tshawb fawb tshuaj tiv thaiv, kev tsim tshuaj thiab lwm yam haujlwm.Nrog rau kev txhim kho biotechnology thiab peptide synthesis thev naus laus zis, ntau thiab ntau cov tshuaj peptide tau tsim thiab siv hauv tsev kho mob.
Muaj ntau yam kev hloov kho peptide, uas tuaj yeem muab faib ua cov kev hloov kho tom qab thiab cov txheej txheem hloov kho (siv cov kev hloov kho amino acid), thiab N-terminal hloov kho, C-terminal hloov kho, sab saw hloov kho, hloov kho amino acid, kev hloov kho pob txha, thiab lwm yam, nyob ntawm qhov chaw hloov kho (Daim duab 1).Raws li ib qho tseem ceeb txhais tau tias hloov lub ntsiab saw hlau los yog sab saw pawg ntawm peptide chains, peptide hloov kho tau zoo hloov lub cev thiab tshuaj zog ntawm peptide tebchaw, ua rau kom dej solubility, ncua lub sij hawm ua hauj lwm nyob rau hauv vivo, hloov lawv lom tis, tshem tawm immunogenicity. , txo cov kev phiv tshuaj lom, thiab lwm yam. Hauv daim ntawv no, ntau lub tswv yim hloov kho peptide loj thiab lawv cov yam ntxwv tau qhia.
1. Cyclization
Cyclic peptides muaj ntau daim ntawv thov hauv biomedicine, thiab ntau lub ntuj peptides nrog kev ua haujlwm lom neeg yog cyclic peptides.Vim hais tias cyclic peptides yuav nruj dua li linear peptides, lawv tsis tshua muaj resistant rau lub plab zom mov, muaj peev xwm ciaj sia nyob rau hauv lub plab zom mov, thiab muaj ib tug muaj zog affinity rau lub hom phiaj receptors.Cyclization yog txoj hauv kev ncaj qha tshaj plaws los ua ke cyclic peptides, tshwj xeeb tshaj yog rau peptides nrog cov pob txha loj.Raws li hom cyclization, nws tuaj yeem muab faib ua sab saw-sab saw hom, davhlau ya nyob twg - sab saw hom, davhlau ya nyob twg - davhlau ya nyob twg hom (kawg mus rau hom kawg).
(1) sidechain-to-sidechain
Feem ntau hom ntawm sab-chain mus rau sab-chain cyclization yog disulfide bridging ntawm cysteine residues.Qhov no cyclization yog qhia los ntawm ib khub ntawm cysteine residues yog deprotected thiab ces oxidized los ua disulfide bonds.Polycyclic synthesis tuaj yeem ua tiav los ntawm kev xaiv tshem tawm ntawm sulfhydryl pawg tiv thaiv.Cyclization yuav ua tau nyob rau hauv ib tug tom qab dissociation hnyav los yog nyob rau hauv ib tug pre-dissociation resin.Cyclization ntawm resins tuaj yeem ua haujlwm tsawg dua li cov kuab tshuaj cyclization vim tias peptides ntawm cov resins tsis tuaj yeem ua rau cyclified conformations.Lwm hom ntawm cov saw-sab-sab saw cyclization yog tsim ntawm cov qauv amide ntawm cov aspartic acid los yog glutamic acid residue thiab cov amino acid hauv paus, uas xav kom cov pab pawg sab saw tiv thaiv yuav tsum muaj peev xwm xaiv tau tshem tawm ntawm polypeptide los yog. ntawm resin los yog tom qab dissociation.Qhov thib peb hom ntawm sab-chain - sab saw cyclization yog tsim ntawm diphenyl ethers los ntawm tyrosine los yog p-hydroxyphenylglycine.Hom cyclization nyob rau hauv cov khoom ntuj tsuas yog pom nyob rau hauv cov khoom microbial, thiab cov khoom cyclization feem ntau muaj peev xwm muaj txiaj ntsig zoo.Kev npaj ntawm cov tebchaw no yuav tsum muaj cov tshuaj tiv thaiv tshwj xeeb, yog li lawv tsis siv ntau zaus hauv kev sib xyaw ntawm cov pa peptides.
(2) Terminal-to-sidechain
Terminal-sab saw cyclization feem ntau suav nrog C-terminal nrog cov amino pawg ntawm lysine lossis ornithine sab saw, lossis N-terminal nrog aspartic acid lossis glutamic acid sab saw.Lwm yam polypeptide cyclization yog tsim los ntawm kev tsim ether bonds ntawm lub davhlau ya nyob twg C thiab serine los yog threonine sab chains.
(3) Terminal los yog lub taub hau-rau-tail hom
Chain polypeptides tuaj yeem raug cycled nyob rau hauv ib qho hnyav los yog tsau rau ntawm ib qho resin los ntawm sab saw cyclation.Tsawg concentrations ntawm peptides yuav tsum tau siv nyob rau hauv hnyav centralization kom tsis txhob oligomerization ntawm peptides.Cov txiaj ntsig ntawm lub taub hau-rau-tail hluavtaws nplhaib polypeptide nyob ntawm qhov sib lawv liag ntawm cov saw polypeptide.Yog li ntawd, ua ntej npaj cyclic peptides rau ntawm qhov loj, ib lub tsev qiv ntawv ntawm qhov ua tau chained lead peptides yuav tsum xub tsim, ua raws li cyclization los nrhiav qhov sib lawv liag nrog cov txiaj ntsig zoo tshaj plaws.
2. N-methylation
N-methylation Ameslikas tshwm sim nyob rau hauv ntuj peptides thiab yog nkag mus rau hauv peptide synthesis los tiv thaiv kev tsim ntawm hydrogen bonds, yog li ua peptides ntau resistant rau biodegradation thiab clearance.Synthesis ntawm peptides siv N-methylated amino acid derivatives yog txoj kev tseem ceeb tshaj plaws.Tsis tas li ntawd, Mitsunobu cov tshuaj tiv thaiv ntawm N-(2-nitrobenzene sulfonyl chloride) polypeptide-resin intermediates nrog methanol kuj siv tau.Txoj kev no tau siv los npaj cov tsev qiv ntawv cyclic peptide uas muaj N-methylated amino acids.
3. Phosphorylation
Phosphorylation yog ib qho kev hloov kho tom qab kev txhais lus ntau tshaj plaws hauv qhov xwm txheej.Hauv tib neeg lub hlwb, ntau dua 30% ntawm cov proteins yog phosphorylated.Phosphorylation, tshwj xeeb tshaj yog thim rov qab phosphorylation, plays lub luag haujlwm tseem ceeb hauv kev tswj ntau cov txheej txheem ntawm tes, xws li cov teeb liab hloov, kev qhia cov noob, lub voj voog ntawm tes thiab kev tswj hwm cytoskeleton, thiab apoptosis.
Phosphorylation tuaj yeem pom ntawm ntau yam amino acid residues, tab sis cov hom phiaj phosphorylation feem ntau yog serine, threonine, thiab tyrosine residues.Phosphotyrosine, phosphothreonine, thiab phosphoserine derivatives tuaj yeem nkag mus rau hauv peptides thaum lub sijhawm ua ke lossis tsim tom qab peptide synthesis.Kev xaiv phosphorylation tuaj yeem ua tiav siv cov seem ntawm serine, threonine, thiab tyrosine uas xaiv tshem tawm cov pab pawg tiv thaiv.Qee cov phosphorylation reagents kuj tuaj yeem qhia phosphoric acid pawg rau hauv polypeptide los ntawm kev hloov kho tom qab.Nyob rau hauv xyoo tas los no, qhov chaw tshwj xeeb phosphorylation ntawm lysine tau ua tiav siv cov tshuaj tiv thaiv Staudinger-phosphite (Daim duab 3).
4. Myristoylation thiab palmitoylation
Acylation ntawm N-terminal nrog fatty acids tso cai rau peptides los yog proteins khi rau cell membranes.Lub myridamoylated ib ntus ntawm N-terminal ua rau Src tsev neeg cov protein kinases thiab thim rov qab transcriptase Gaq proteins kom raug tsom mus khi rau cov cell membranes.Myristic acid tau txuas mus rau N-terminal ntawm resin-polypeptide siv cov txheej txheem sib txuas, thiab cov lipopeptide tshwm sim tuaj yeem raug cais tawm raws li cov qauv txheej txheem thiab ua kom huv los ntawm RP-HPLC.
5. Glycosylation
Glycopeptides xws li vancomycin thiab teicolanin yog cov tshuaj tua kab mob tseem ceeb rau kev kho cov kab mob tiv thaiv kab mob, thiab lwm yam glycopeptides feem ntau siv los txhawb kev tiv thaiv kab mob.Tsis tas li ntawd, txij li ntau cov kab mob microbial antigens yog glycosylated, nws yog ib qho tseem ceeb heev los kawm glycopeptides rau kev txhim kho cov kev kho mob ntawm tus kab mob.Ntawm qhov tod tes, nws tau pom tias cov proteins ntawm cov cell membrane ntawm cov qog hlwb pom cov glycosylation txawv txav, uas ua rau glycopeptides ua lub luag haujlwm tseem ceeb hauv kev tshawb nrhiav qog nqaij hlav thiab tiv thaiv qog noj ntshav.Glycopeptides yog npaj los ntawm Fmoc/t-Bu txoj kev.Glycosylated residues, xws li threonine thiab serine, feem ntau nkag mus rau hauv polypeptides los ntawm pentafluorophenol ester activated fMOCs los tiv thaiv glycosylated amino acids.
6. Isoprene
Isopentadienylation tshwm sim ntawm cysteine residues nyob rau hauv lub sab saw ze ntawm C-terminal.Protein isoprene tuaj yeem txhim kho cell membrane affinity thiab tsim cov protein-protein sib cuam tshuam.Isopentadienated proteins muaj xws li tyrosine phosphatase, me GTase, cochaperone molecules, nuclear lamina, thiab centromeric binding proteins.Isoprene polypeptides tuaj yeem npaj siv isoprene ntawm resins lossis los ntawm kev qhia txog cysteine derivatives.
7. Polyethylene glycol (PEG) kev hloov kho
Kev hloov kho PEG tuaj yeem siv los txhim kho cov protein hydrolytic stability, biodistribution thiab peptide solubility.Kev taw qhia ntawm PEG chains rau peptides tuaj yeem txhim kho lawv cov tshuaj pharmacological thiab tseem inhibit hydrolysis ntawm peptides los ntawm proteolytic enzymes.PEG peptides dhau los ntawm glomerular capillary hla ntu yooj yim dua li cov peptides zoo tib yam, txo qis lub raum tshem tawm.Vim qhov kev ncua ntev ib nrab-lub neej ntawm PEG peptides hauv vivo, qib kev kho mob ib txwm tuaj yeem tswj tau nrog cov koob tshuaj qis dua thiab tsis tshua muaj tshuaj peptide.Txawm li cas los xij, kev hloov kho PEG kuj muaj qhov tsis zoo.Cov nyiaj ntau ntawm PEG tiv thaiv cov enzyme los ntawm degrading peptide thiab kuj txo cov peptide txoj kev khi rau lub hom phiaj receptor.Tab sis PEG peptides 'kev sib raug zoo tsis tshua muaj feem ntau cuam tshuam los ntawm lawv cov tshuaj pharmacokinetic ib nrab-lub neej ntev, thiab los ntawm kev muaj nyob hauv lub cev ntev dua, PEG peptides muaj qhov zoo dua ntawm kev nqus mus rau hauv cov ntaub so ntswg.Yog li ntawd, PEG polymer specifications yuav tsum tau optimized rau kev pom zoo.Ntawm qhov tod tes, PEG peptides sib sau ua ke hauv daim siab vim txo lub raum tshem tawm, ua rau macromolecular syndrome.Yog li ntawd, kev hloov kho PEG yuav tsum tau tsim kom zoo dua qub thaum siv peptides rau kev sim tshuaj.
Kev hloov pauv ntau pawg ntawm PEG modifiers tuaj yeem raug sau tseg raws li hauv qab no: 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 (xws li propional-ald, butyrALD), acrylic puag (-acrylate-acrl), azido-azide, Biotin, Fluorescein, glutaryl -GA, Acrylate Hydrazide, alkyne-alkyne, p-toluenesulfonate -OTs, succinimide succinate -SS, thiab lwm yam. PEG derivatives nrog carboxylic acids tuaj yeem ua ke rau n-terminal amines lossis lysine sides.Amino-activated PEG tuaj yeem ua ke rau aspartic acid lossis glutamic acid sab chains.Mal-activated PEG tuaj yeem txuas rau mercaptan ntawm tag nrho kev tiv thaiv cysteine sab chains [11].PEG modifiers feem ntau cais raws li hauv qab no (ceeb toom: mPEG yog methoxy-PEG, CH3O-(CH2CH2O)n-CH2CH2-OH):
(1) Ncaj saw PEG hloov kho
mPEG-SC, mPEG-SCM, mPEG-SPA, mPEG-OTs, mPEG-SH, mPEG-ALD, mPEG-butyrALD, mPEG-SS
(2) bifunctional PEG hloov kho
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 tuaj yeem ua tau ruaj khov nrog avidin lossis streptavidin, thiab lub zog ruaj khov txawm nyob ze rau covalent daim ntawv cog lus.Biotin-labeled peptides feem ntau yog siv hauv immunoassay, histocytochemistry, thiab fluorescence-based flow cytometry.Labeled antibiotin cov tshuaj tiv thaiv kuj tuaj yeem siv los khi biotinylated peptides.Biotin cov ntawv lo feem ntau txuas rau lysine sab saw lossis N davhlau ya nyob twg.6-aminocaproic acid feem ntau yog siv los ua kev sib txuas ntawm peptides thiab biotin.Daim ntawv cog lus yog hloov tau yooj yim hauv kev khi rau lub substrate thiab khi zoo dua nyob rau hauv lub xub ntiag ntawm steric hindrance.
9. Fluorescent labeling
Fluorescent labeling tuaj yeem siv los taug qab polypeptides hauv cov hlwb nyob thiab kawm txog cov enzymes thiab cov txheej txheem ntawm kev ua.Tryptophan (Trp) yog cov fluorescent, yog li nws tuaj yeem siv rau kev sau npe hauv qab.Lub emission spectrum ntawm tryptophan nyob ntawm lub peripheral ib puag ncig thiab txo qis nrog cov kuab tshuaj polarity tsawg, cov cuab yeej uas muaj txiaj ntsig zoo rau kev kuaj peptide qauv thiab receptor khi.Tryptophan fluorescence tuaj yeem quenched los ntawm protonated aspartic acid thiab glutamic acid, uas yuav txwv tsis pub siv.Dansyl chloride pawg (Dansyl) yog cov fluorescent heev thaum khi rau ib pawg amino thiab feem ntau yog siv los ua ib daim ntawv lo rau cov amino acids lossis cov proteins.
Fluorescence resonance Zog hloov dua siab tshiab (FRET) muaj txiaj ntsig zoo rau kev tshawb fawb enzyme.Thaum siv FRET, lub substrate polypeptide feem ntau muaj cov fluorescence-labeling pawg thiab ib pawg fluorescence-quenching.Cov npe fluorescent pawg yog quenched los ntawm quencher los ntawm tsis-photon zog hloov.Thaum cov peptide dissociated los ntawm cov enzyme nyob rau hauv nqe lus nug, lub labeling pab pawg neeg emits fluorescence.
10. Cage polypeptides
Cage peptides muaj optically tshem tau cov pab pawg tiv thaiv uas thaiv cov peptide los ntawm kev khi rau cov receptor.Thaum raug UV hluav taws xob, peptide tau qhib, rov ua kom nws txoj kev sib raug zoo rau cov receptor.Vim tias qhov kev ua kom pom qhov muag no tuaj yeem tswj tau raws li lub sijhawm, amplitude, lossis qhov chaw, cage peptides tuaj yeem siv los kawm txog kev cuam tshuam tshwm sim hauv hlwb.Cov pab pawg tiv thaiv feem ntau siv rau lub tawb polypeptides yog 2-nitrobenzyl pawg thiab lawv cov derivatives, uas tuaj yeem nkag rau hauv peptide synthesis ntawm kev tiv thaiv amino acid derivatives.Amino acid derivatives uas tau tsim yog lysine, cysteine, serine, thiab tyrosine.Aspartate thiab glutamate derivatives, txawm li cas los xij, tsis tau siv ntau vim lawv raug rau cyclization thaum peptide synthesis thiab dissociation.
11. Polyantigenic peptide (MAP)
Cov peptides luv luv feem ntau tsis muaj kev tiv thaiv kab mob thiab yuav tsum tau ua ke rau cov kab mob cab los tsim cov tshuaj tiv thaiv.Polyantigenic peptide (MAP) yog tsim los ntawm ntau yam peptides zoo tib yam txuas nrog lysine nuclei, uas tuaj yeem qhia tshwj xeeb muaj zog tiv thaiv kab mob thiab tuaj yeem siv los npaj peptide-carrier protein couplets.MAP polypeptides tuaj yeem ua ke los ntawm cov khoom sib txuas ntawm MAP resin.Txawm li cas los xij, kev sib txuas tsis tiav ua rau ploj lossis raug txiav peptide chains ntawm qee ceg thiab yog li tsis pom cov khoom ntawm thawj MAP polypeptide.Raws li lwm txoj hauv kev, peptides tuaj yeem npaj thiab huv si sib cais thiab tom qab ntawd ua ke rau MAP.Cov peptide ib ntus txuas nrog peptide core yog qhov zoo-txhais thiab yooj yim yam ntxwv los ntawm huab hwm coj spectrometry.
Xaus
Kev hloov pauv Peptide yog ib qho tseem ceeb ntawm kev tsim peptides.Cov tshuaj hloov kho peptides tuaj yeem tsis tsuas yog tswj hwm kev ua haujlwm lom neeg siab, tab sis kuj tseem tiv thaiv qhov tsis zoo ntawm kev tiv thaiv kab mob thiab tshuaj lom.Nyob rau tib lub sijhawm, kev hloov kho tshuaj lom neeg tuaj yeem ua rau peptides nrog qee cov khoom zoo tshiab.Nyob rau hauv xyoo tas los no, txoj kev ntawm CH ua kom muaj kev hloov kho tom qab ntawm polypeptides tau tsim sai heev, thiab ntau qhov tseem ceeb tau ua tiav.
Post lub sij hawm: Mar-20-2023