Ma peptides ndi gulu lamagulu opangidwa ndi kulumikizana kwa ma amino acid angapo kudzera m'magulu a peptide.Amapezeka paliponse m'zamoyo.Mpaka pano, ma peptides zikwi makumi ambiri apezeka m'zamoyo.Ma peptides amagwira ntchito yofunikira pakuwongolera magwiridwe antchito a machitidwe osiyanasiyana, ziwalo, minyewa ndi ma cell komanso zochitika zamoyo, ndipo nthawi zambiri amagwiritsidwa ntchito pakuwunika magwiridwe antchito, kafukufuku wa antibody, chitukuko cha mankhwala ndi zina.Ndi chitukuko cha biotechnology ndi peptide synthesis, mankhwala ochulukirapo a peptide apangidwa ndikugwiritsidwa ntchito kuchipatala.
Pali mitundu yosiyanasiyana ya peptide zosinthidwa, zomwe zingagawidwe mophweka positi ndi kusintha ndondomeko (pogwiritsa ntchito kusinthidwa kwa amino acid), ndi kusinthidwa kwa N-terminal, kusinthidwa kwa C-terminal, kusinthidwa kwa unyolo, amino acid kusinthidwa, kusinthidwa kwa mafupa, etc., kutengera malo osinthidwa (Chithunzi 1).Monga njira yofunikira yosinthira unyolo waukulu kapena magulu am'mbali a unyolo wa peptide, kusinthidwa kwa peptide kumatha kusintha bwino mawonekedwe a thupi ndi mankhwala amafuta a peptide, kuonjezera kusungunuka kwamadzi, kutalikitsa nthawi yakuchita mu vivo, kusintha kugawa kwawo kwachilengedwe, kuthetsa immunogenicity. , kuchepetsa zotsatira za poizoni, ndi zina zotero.
1. Kuyenda panjinga
Ma cyclic peptides ali ndi ntchito zambiri mu biomedicine, ndipo ma peptides ambiri achilengedwe okhala ndi biology ndi ma cyclic peptides.Chifukwa ma cyclic peptides amakhala olimba kwambiri kuposa ma peptide am'mbali, amalimbana kwambiri ndi kugaya chakudya, amatha kukhala ndi moyo m'mimba, ndikuwonetsa kuyanjana kolimba kwa zolandilira.Cyclization ndiye njira yolunjika kwambiri yopangira ma cyclic peptides, makamaka ma peptides okhala ndi mafupa akulu omangika.Malinga ndi ma cyclization mode, imatha kugawidwa m'magulu am'mbali mwa unyolo, mtundu wa unyolo wam'mbali, terminal - terminal mtundu (mapeto mpaka kumapeto).
(1) sidechain-to-sidechain
Mtundu wodziwika bwino wa unyolo wam'mbali kupita ku unyolo wam'mbali ndi kulumikiza kwa disulfide pakati pa zotsalira za cysteine.Kuyendetsa uku kumayambitsidwa ndi zotsalira za cysteine zotetezedwa ndikusinthidwa kukhala zomangira za disulfide.Kuphatikizika kwa polycyclic kumatha kutheka pochotsa magulu achitetezo a sulfhydryl.Kuyendetsa njinga kumatha kuchitika mu chosungunulira cha post-dissociation kapena pa pre-dissociation resin.Kuthamanga kwa ma resins kungakhale kocheperako poyerekeza ndi zosungunulira zosungunulira chifukwa ma peptides pa resin sapanga ma cyclified conformations.Mtundu wina wa unyolo wam'mbali - cyclization yam'mbali ndi mapangidwe amipangidwe amide pakati pa aspartic acid kapena zotsalira za glutamic acid ndi maziko a amino acid, zomwe zimafuna kuti gulu lachitetezo chakumbali lizitha kuchotsedwa mwa kusankha ku polypeptide mwina. pa utomoni kapena pambuyo dissociation.Mtundu wachitatu wa unyolo wam'mbali - cyclization yam'mbali ndi mapangidwe a diphenyl ethers ndi tyrosine kapena p-hydroxyphenylglycine.Kuthamanga kwamtundu wotere muzinthu zachilengedwe kumapezeka muzinthu zazing'ono zokha, ndipo zopangira ma cyclization nthawi zambiri zimakhala ndi mankhwala.Kukonzekera kwa mankhwalawa kumafuna zinthu zapadera zomwe zimachitika, kotero sizimagwiritsidwa ntchito popanga ma peptides wamba.
(2) terminal-to-sidechain
Kuthamanga kwa unyolo wam'mbali nthawi zambiri kumaphatikizapo C-terminal ndi gulu la amino la lysine kapena ornithine, kapena N-terminal yokhala ndi aspartic acid kapena glutamic acid.Kuzungulira kwina kwa polypeptide kumapangidwa ndikupanga zomangira za ether pakati pa terminal C ndi unyolo wam'mbali wa serine kapena threonine.
(3) Mtundu wa terminal kapena mutu ndi mchira
Unyolo polypeptides akhoza mwina panjinga mu zosungunulira kapena kuikidwa pa utomoni ndi mbali unyolo cyclation.Ma peptides otsika ayenera kugwiritsidwa ntchito posungunulira centralization kupewa oligomerization ya peptides.Zokolola za polypeptide yopangidwa ndi mutu-to-mchira zimatengera kutsata kwa unyolo wa polypeptide.Chifukwa chake, musanakonzekere ma cyclic peptides pamlingo waukulu, laibulale ya ma peptide otsogola omwe ali ndi unyolo iyenera kupangidwa kaye, ndikutsatiridwa ndi cyclization kuti mupeze zotsatira zake zabwino.
2. N-methylation
N-methylation poyambilira imapezeka mu peptide yachilengedwe ndipo imalowetsedwa mu kaphatikizidwe ka peptide kuti ateteze mapangidwe a haidrojeni zomangira, potero zimapangitsa ma peptides kukhala osagwirizana ndi biodegradation ndi chilolezo.Kaphatikizidwe ka peptides pogwiritsa ntchito zotumphukira za N-methylated amino acid ndiyo njira yofunika kwambiri.Kuphatikiza apo, Mitsunobu reaction ya N-(2-nitrobenzene sulfonyl chloride) polypeptide-resin intermediates ndi methanol ingagwiritsidwenso ntchito.Njirayi yagwiritsidwa ntchito pokonzekera malaibulale a cyclic peptide okhala ndi N-methylated amino acid.
3. Phosphorylation
Phosphorylation ndi imodzi mwazosintha zomwe zimachitika pambuyo pomasulira m'chilengedwe.M'maselo aumunthu, mapuloteni opitilira 30% amakhala ndi phosphorylated.Phosphorylation, makamaka phosphorylation yosinthika, imakhala ndi gawo lofunikira pakuwongolera njira zambiri zama cell, monga kusintha kwa ma sign, gene expression, cell cycle and cytoskeleton regulation, and apoptosis.
Phosphorylation imatha kuwonedwa pazotsalira zosiyanasiyana za amino acid, koma zotsalira za phosphorylation zodziwika bwino ndi serine, threonine, ndi zotsalira za tyrosine.Phosphotyrosine, phosphothreonine, ndi zotumphukira za phosphoserine zitha kulowetsedwa mu peptides panthawi ya kaphatikizidwe kapena kupangidwa pambuyo pa kaphatikizidwe ka peptide.Kusankha phosphorylation kumatha kupezedwa pogwiritsa ntchito zotsalira za serine, threonine, ndi tyrosine zomwe zimachotsa magulu oteteza.Ma reagents ena a phosphorylation amathanso kuyambitsa magulu a phosphoric acid mu polypeptide posintha positi.M'zaka zaposachedwa, phosphorylation yeniyeni ya lysine yakhala ikugwiritsidwa ntchito pogwiritsa ntchito mankhwala a Staudinger-phosphite reaction (Chithunzi 3).
4. Myristoylation ndi palmitoylation
Acylation ya N-terminal yokhala ndi mafuta acids imalola ma peptides kapena mapuloteni kumangirira ku nembanemba yama cell.Kutsatizana kwa myridamoylated pa N-terminal kumathandizira kuti mapuloteni a Src a banja la Src ndi reverse transcriptase Gaq agwirizane ndi ma cell.Myristic acid idalumikizidwa ndi N-terminal ya resin-polypeptide pogwiritsa ntchito njira zolumikizirana, ndipo zotsatira zake za lipopeptide zitha kulekanitsidwa pamikhalidwe yokhazikika ndikuyeretsedwa ndi RP-HPLC.
5. Glycosylation
Glycopeptides monga vancomycin ndi teicolanin ndi maantibayotiki ofunikira pochiza matenda a bakiteriya osamva mankhwala, ndipo ma glycopeptides ena nthawi zambiri amagwiritsidwa ntchito kulimbikitsa chitetezo chamthupi.Kuphatikiza apo, popeza ma antigen ambiri amapangidwa ndi glycosylated, ndikofunikira kwambiri kuphunzira ma glycopeptides kuti apititse patsogolo machiritso a matenda.Kumbali inayi, zapezeka kuti mapuloteni omwe ali pa cell membrane yama cell chotupa amawonetsa glycosylation, zomwe zimapangitsa kuti glycopeptides itenge gawo lofunikira pakufufuza kwa khansa komanso chitetezo chamthupi chotupa.Glycopeptides amakonzedwa ndi njira ya Fmoc/t-Bu.Zotsalira za glycosylated, monga threonine ndi serine, nthawi zambiri zimalowetsedwa mu polypeptides ndi pentafluorophenol ester activated fMOCs kuteteza glycosylated amino acid.
6. Isoprene
Isopentadienylation imapezeka pa zotsalira za cysteine mu unyolo wam'mbali pafupi ndi C-terminal.Mapuloteni a isoprene amatha kusintha kulumikizana kwa membrane wa cell ndikupanga kulumikizana kwa mapuloteni ndi mapuloteni.Mapuloteni omwe ali ndi Isopentadienate amaphatikizapo tyrosine phosphatase, GTase yaying'ono, mamolekyu a cochaperone, lamina ya nyukiliya, ndi mapuloteni omangira centromeric.Isoprene polypeptides ikhoza kukonzedwa pogwiritsa ntchito isoprene pa resins kapena poyambitsa zotumphukira za cysteine.
7. Kusintha kwa polyethylene glycol (PEG).
Kusintha kwa PEG kungagwiritsidwe ntchito kukonza kukhazikika kwa protein hydrolytic, biodistribution ndi kusungunuka kwa peptide.Kuyambitsidwa kwa unyolo wa PEG ku ma peptides kumatha kusintha mawonekedwe awo azachipatala komanso kuletsa hydrolysis ya peptides ndi michere ya proteinolytic.PEG peptides amadutsa mu glomerular capillary cross section mosavuta kuposa ma peptide wamba, amachepetsa kwambiri kuchotsedwa kwa aimpso.Chifukwa chotalikirapo theka la moyo wa PEG peptides mu vivo, mulingo wabwinobwino wamankhwala ukhoza kusungidwa ndi Mlingo wocheperako komanso mankhwala ochepera a peptide.Komabe, kusinthidwa kwa PEG kumakhalanso ndi zotsatirapo zoipa.Kuchuluka kwa PEG kumalepheretsa enzyme kuti isawononge peptide komanso kumachepetsa kumangirira kwa peptide ku cholandirira chomwe mukufuna.Koma kuyanjana kochepa kwa PEG peptides nthawi zambiri kumathetsedwa ndi theka la moyo wawo wautali wa pharmacokinetic, ndipo pokhalapo m'thupi nthawi yayitali, ma peptide a PEG amakhala ndi mwayi wochuluka wolowetsedwa mu minofu yomwe mukufuna.Chifukwa chake, mawonekedwe a PEG polima akuyenera kukonzedwa kuti akhale ndi zotsatira zabwino.Kumbali ina, ma peptides a PEG amawunjikana m'chiwindi chifukwa cha kuchepa kwa aimpso, zomwe zimapangitsa kuti pakhale macromolecular syndrome.Choncho, kusintha kwa PEG kuyenera kupangidwa mosamala kwambiri pamene ma peptide amagwiritsidwa ntchito poyesa mankhwala.
Magulu osinthika a PEG modifiers akhoza kufotokozedwa mwachidule motere: 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 (monga propional-ald, butyrALD), acrylic base (-acrylate-acrl), azido-azide, biotinyl - Biotin, Fluorescein, glutaryl -GA, Acrylate Hydrazide, alkyne-alkyne, p-toluenesulfonate -OTs, succinimide succinate -SS, etc. PEG zotumphukira ndi carboxylic zidulo akhoza kuphatikizidwa n-terminal amines kapena lysine mbali unyolo.Amino-activated PEG imatha kuphatikizidwa ndi aspartic acid kapena glutamic acid side chain.PEG yokhala ndi mal-activated imatha kulumikizidwa ku mercaptan ya unyolo wam'mbali wa cysteine wotetezedwa [11].Zosintha za PEG zimagawidwa motere (zindikirani: mPEG ndi methoxy-PEG, CH3O-(CH2CH2O)n-CH2CH2-OH) :
(1) chosinthira cha PEG chowongoka
mPEG-SC, mPEG-SCM, mPEG-SPA, mPEG-OTs, mPEG-SH, mPEG-ALD, mPEG-butyrALD, mPEG-SS
(2) kusintha kwa PEG kosagwira ntchito
HCOO-PEG-COOH, NH2-PEG-NH2, OH-PEG-COOH, OH-PEG-NH2, HCl·NH2-PEG-COOH, MAL-PEG-NHS
(3) nthambi ya PEG modifier
(mPEG)2-NHS, (mPEG)2-ALD, (mPEG)2-NH2, (mPEG)2-MAL
8. Biotinization
Biotin akhoza kumangidwa mwamphamvu ndi avidin kapena streptavidin, ndipo mphamvu yomangiriza imakhala pafupi ndi mgwirizano wa covalent.Ma peptides olembedwa ndi biotin amagwiritsidwa ntchito kwambiri mu immunoassay, histocytochemistry, ndi fluorescence-based flow cytometry.Ma antibodies otchedwa antibiotin amatha kugwiritsidwanso ntchito kumanga ma peptide a biotinylated.Zolemba za Biotin nthawi zambiri zimamangiriridwa ku tcheni cham'mbali cha lysine kapena N terminal.6-aminocaproic acid nthawi zambiri amagwiritsidwa ntchito ngati chomangira pakati pa peptides ndi biotin.Chomangiracho chimasinthasintha pomangiriza ku gawo lapansi ndikumangirira bwino pamaso pa chopinga cha steric.
9. Kulemba kwa fluorescent
Kulemba kwa fluorescent kumatha kugwiritsidwa ntchito kutsata ma polypeptides m'maselo amoyo komanso kuphunzira ma enzyme ndi njira zogwirira ntchito.Tryptophan (Trp) ndi fulorosenti, choncho itha kugwiritsidwa ntchito polemba zilembo zamkati.Kutulutsa kwa tryptophan kumadalira malo am'mphepete mwake ndipo kumachepa ndi kuchepa kwa zosungunulira polarity, chinthu chomwe chimakhala chothandiza pozindikira mawonekedwe a peptide ndi kumanga kwa ma receptor.Tryptophan fluorescence imatha kuzimitsidwa ndi protonated aspartic acid ndi glutamic acid, zomwe zingachepetse kugwiritsidwa ntchito kwake.Gulu la Dansyl chloride (Dansyl) limapangidwa ndi fulorosenti kwambiri likamangika ku gulu la amino ndipo nthawi zambiri limagwiritsidwa ntchito ngati chizindikiro cha fulorosenti ya ma amino acid kapena mapuloteni.
Fluorescence resonance Energy kutembenuka (FRET) ndiyothandiza pamaphunziro a enzyme.FRET ikagwiritsidwa ntchito, gawo lapansi la polypeptide nthawi zambiri limakhala ndi gulu la fluorescence-labeling ndi gulu la fluorescence-quenching.Magulu olembedwa a fulorosenti amazimitsidwa ndi chozimitsira pogwiritsa ntchito mphamvu zopanda photon.Peptide ikasiyanitsidwa ndi enzyme yomwe ikufunsidwa, gulu lolemba limatulutsa fluorescence.
10. Ma polypeptides a khola
Ma Cage peptides ali ndi magulu oteteza omwe amatha kuchotsedwa omwe amatchinjiriza peptide kuti isamangike ndi cholandirira.Ikawonetsedwa ndi kuwala kwa UV, peptide imayatsidwa, ndikubwezeretsa kuyanjana kwake ndi cholandirira.Chifukwa kutsegulira kwa kuwalaku kumatha kuyendetsedwa molingana ndi nthawi, matalikidwe, kapena malo, ma peptides a khola angagwiritsidwe ntchito pophunzira zomwe zimachitika m'maselo.Magulu omwe amagwiritsidwa ntchito kwambiri poteteza khola la polypeptides ndi magulu a 2-nitrobenzyl ndi zotumphukira zake, zomwe zitha kuyambitsidwa mu kaphatikizidwe ka peptide kudzera mwa zotumphukira za amino acid.Zochokera ku amino acid zomwe zapangidwa ndi lysine, cysteine, serine, ndi tyrosine.Zotumphukira za aspartate ndi glutamate, komabe, sizimagwiritsidwa ntchito nthawi zambiri chifukwa chotengeka ndi ma cyclization panthawi ya peptide synthesis and dissociation.
11. Polyantigenic peptide (MAP)
Ma peptide amfupi nthawi zambiri satetezedwa ndipo amayenera kuphatikizidwa ndi mapuloteni onyamula kuti apange ma antibodies.Polyantigenic peptide (MAP) imapangidwa ndi ma peptide angapo ofanana olumikizidwa ndi lysine nuclei, omwe amatha kufotokoza mwachindunji ma immunogens amphamvu kwambiri ndipo angagwiritsidwe ntchito popanga ma peptide-carrier protein couplets.MAP ma polypeptides amatha kupangidwa ndi kaphatikizidwe kolimba kagawo pa MAP resin.Komabe, kulumikizana kosakwanira kumabweretsa unyolo wa peptide wosowa kapena wocheperako panthambi zina motero siziwonetsa mawonekedwe a MAP polypeptide yoyambirira.M'malo mwake, ma peptides amatha kukonzedwa ndikuyeretsedwa padera ndikuphatikizidwa ku MAP.Kutsatizana kwa peptide komwe kumalumikizidwa pachimake cha peptide kumatanthauzidwa bwino komanso kumadziwika mosavuta ndi misa spectrometry.
Mapeto
Kusintha kwa peptide ndi njira yofunikira yopangira ma peptides.Mankhwala kusinthidwa peptides sangakhoze kukhalabe mkulu kwachilengedwenso ntchito, komanso mogwira kupewa zovuta za immunogenicity ndi kawopsedwe.Nthawi yomweyo, kusinthidwa kwamankhwala kumatha kupatsa ma peptides ndi zinthu zina zabwino kwambiri.M'zaka zaposachedwa, njira ya CH activation for post-modified of polypeptides yapangidwa mofulumira, ndipo zotsatira zambiri zofunika zakwaniritsidwa.
Nthawi yotumiza: Mar-20-2023