2019 Novel Coronavirus SARS-CoV-2 is a virus identified as the cause of an outbreak of respiratory illness Covid-19 first detected in Wuhan, China.
To help expedite Covid-19 research, LifeTein synthesized a 69 amino acid spike glycoprotein with one disulfide bond in 6 days. This effort is a partnership with a biotech company for drug development.
Cyclic peptides as broad-spectrum antiviral agents
Antiviral drugs and vaccines are the most powerful tools to combat viral diseases. Most drugs and vaccines only target a single virus. However, the broad-spectrum antivirals can be used for rapid management of new or drug-resistant viral strains. Cyclized peptides and peptide analogs are excellent examples of broad-spectrum antivirals.
An artificial peptide molecule was found to neutralize a broad range of group 1 influenza A viruses, including H5N1. The peptide design was based on complementarity determining region (CDR) loops have been reported for other viral targets. The optimized peptides bind to the highly conserved stem epitope and block the low pH-induced conformational rearrangements associated with membrane fusion.
These peptidic compounds and their advantageous biological properties should accelerate development of novel small molecule and peptide-based therapeutics against influenza virus.
The linear peptide is Suc-SQLRSLEYFEWLSQ-NH2. Three cyclization strategies were used: head to tail, side chain to side chain and side chain to tail. An ornithine (Orn) side chain was fused with the carboxyl terminus of β-alanine for lactam formation.
Check here for more details: Potent peptidic fusion inhibitors of influenza virus, Science 28 Sep 2017, DOI: 10.1126/science.aan0516
Lately, more broad-spectrum antiviral agents were found to target viruses. It was found that 55 compounds can target eight different RNA and DNA viruses. Dalbavancin is a novel lipo-glycopeptide antibiotic. The lipoglycopeptide disrupts bacterial cell wall formation by binding to
the terminal d-alanyl-d-alanine peptidoglycan sequence in Gram-positive bacteria in a linear, concentration-dependent manner. The dalbavancin has effects on echovirus 1, ezetimibe against HIV1 and Zika virus.
More details: https://www.ncbi.nlm.nih.gov/pubmed/29698664
Protein Purification Using Magnetic Beads: Top Tips for Success
Magnetic bead-based protein purification offers a powerful solution for various applications like high-throughput microscale purification, pull-down/CoIP experiments, and protein-protein or protein-DNA interaction studies. Here’s why magnetic beads are the top choice: they can be coated with specific affinity ligands for antigens, antibodies, proteins, or nucleic acids. Moreover, magnetic beads are non-porous and have a defined diameter, eliminating hidden surfaces where molecules can stick, leading to reduced background, simplified purification, and streamlined washing steps. Compared to traditional bead separation methods involving agarose, sepharose, or silica beads, magnetic bead separation stands out as the quickest, cleanest, and most efficient technique.
If you’re new to working with magnetic beads, here are some essential tips to ensure success:
By following these tips, you can make the most of magnetic bead-based protein purification, improving the efficiency and reliability of your experiments.
Giantin, a novel conserved Golgi membrane protein, is a disulfide-linked homodimer. It was found that BFA-induced Golgi disorganization is associated with the monomerization of giantin.
The pull-down experiment was performed. The control peptide biotin-GHGTGSTGSGSMLRTLLRRRL synthesized by LifeTein was incubated with lysate and Dynabeads, as well as the lysate incubated with Dynabeads only served as a control. Dynabeads carrying MGAT1 peptide were able to pull-down giantin from the lysate of HeLa cells, however, giantin was not detected in the pull-down fraction from the lysate exposed to the Dynabeads or in combination with control peptide. It is logical to hypothesize that the MGAT1 binding domain of giantin lies within its N-terminal non-coiled-coil area.
The Dynabeads function similarly to LifeTein magnetic beads:
https://www.lifetein.com/peptide-product/amineactivated-peptide-conjugation-magnetic-beads-p-3647.html
Cells 2019, 8(12), 1631; https://doi.org/10.3390/cells8121631
Table 1 Selection of cell-penetrating peptides
(Reference: https://doi.org/10.1007/978-981-13-8747-0, Ülo Langel, CPP, Cell-Penetrating Peptides, 2019)
| Name | Sequence |
| 2 | DSLKSYWYLQKFSWR (Kondo et al. 2012) |
| 18A | DWLKAFYDKVAEKLKEAF (Datta et al. 2000) |
| α1H | KSKTEYYNAWAVWERNAP (Gomarasca et al. 2017) |
| α2H | GNGEQREMAVSRLRDCLDRQA (Gomarasca et al. 2017) |
| A22p | HTPGNSNKWKHLQENKKGRPRR (Shin et al. 2014) |
| Ac-18A-NH2 | DWLKAFYDKVAEKLKEAF) (Wimley and White 2000) |
| aCPP | Typical sequence R9GPLGLAGE8 (Li et al 2015) |
| AdVpVI(33-55) | Ac-GAFSWGSLWSGIKNFGSTVKNYG (Murayama et al. 2016) |
| AIP6 | RLRWR (Wang et al. 2011) |
| all-d DsC18 | Glrkrlrkfrnkikek (Bergmann et al. 2017) |
| αgliadin(31-43) | LGQQQPFPPQQPY (Paolella et al. 2018) |
| Alyteserin-2a | ILGKLLSTAAGLLSNL (Conlon et al. 2013) |
| ANG | TFFYGGSRGKRNNFKTEEY (Demeule et al. 2008) |
| ApoE(141–150) | Ac-LRKLRKRLLRX-Bpg-G (Shabanpoor et al. 2017) |
| ApoE-derived | Ac-LRKLRKRLLR (Tailhades et al. 2017) |
| Arf(1-22) | MVRRFLVTLRIRRACGPPRVRV (Johansson et al. 2008) |
| AT1002 | FCIGRL (Gopalakrishnan et al. 2009) |
| AT1AR(304-318) | FLGKKFKKYFLQLLK (Östlund et al. 2005) |
| Bac7 | RRIRPRPPRLPRPRPRPLPFPRPGPRPIPRPL (Sadler et al. 2002) |
| BGPC7-FHV | RRRRNRTRRNRRRVR-RRFYGPV (Wongso et al. 2017) |
| Bim | EIWIAQELRRIGDEFNAYYARLLC (Kim et al. 2017) |
| BP16 | KKLFKKILKKL (Soler et al. 2014) |
| BP100 | KKLFKKILKYL (Eggenberger et al. 2009) |
| BPP13a | GGWPRPGPEIPP (Sciani et al. 2017) |
| bPrPp(1-30) | MVKSKIGSWILVLFVAMWSDVGLCKKRPKP (Magzoub et al. 2006) |
| BR2 | RAGLQFPVGRLLRRLLR (Lim et al. 2013) |
| Buforin II | TRSSRAGLQFPVGRVIIRLLRK (Park et al.1998) |
| Buforin IIb | RAGLQFPVG[RLLR]3 (Lee et al. 2008) |
| C6M1 | RLWRLLWRLWRRLWRLLR (Jafari et al. 2014) |
| C105Y | CSIPPEVKFNKPFVYLI (Rhee and Davis 2006) |
| CADY | GLWRALWRLLRSLWRLLWRA cycteamide (Crombez et al. 2009a) |
| CAR | CARSKNKDC (Toba et al. 2014) |
| CA-Tat | KWKLFKKYGRKKRRQRRR (Lv et al. 2017) |
| CB5005 M | KLKLALALALA (Zhang et al. 2016) |
| CDB3 | REDEDEIEW (Issaeva et al. 2003) |
| CendR | RPARPAR (Hu et al. 2014) |
| cF<tR4 | cyclic F<tRRRRQ (Qian et al. 2014) |
| CGKRK | CGKRK (Griffin et al. 2017) |
| CIGB-300 | cyclic CWMSPRHLGTC-Tat (Perera et al. 2012) |
| CIGB-552 | Ac-HARIKpTFRRlKWKYKGKFW (Fernandez Masso et al. 2013) |
| CLIP6 | KVRVRVRVpPTRVRERVK (Soudah et al. 2017) |
| CooP | ACGLSGLGVA (Hyvonen et al. 2014) |
| CpMTP | ARLLWLLRGLTLGTAPRRA (Jain and Chugh 2016) |
| CPNT | STSGTGKMTRAQRRAAARRNRA (Qi et al. 2011) |
| CPP1 | (KFF)3K (Patel et al. 2017) |
| CPP33 | RLWMRWYSPRTRAYG (Lin et al. 2018) |
| CPP-C | PIEVCMYREP (Nakayama et al. 2011) |
| CPPecp | NYRWRCKNQN (Fu et al. 2017) |
| C-peptide | GPGLWERQAREHSERKKRRRESECKAA (Fan et al. 2016) |
| CRGDK | CRGDK (Zhao et al. 2018) |
| Crotamine | YKQCHKKGGHCFPKEKICLPPSSDFGKMDCRWRWKCCKKGSG (Rodrigues et al. 2012) |
| cSN50 | AAVALLPAVLLALLAPVQRKRQKLMP (Torgerson et al. 1998) |
| 65-2CTS | CPYVNQRPQKARYRNG (Percipalle et al. 2003) |
| CWR8K | CWR8K (Sasaki et al. 2008) |
| CyLoP-1 | CRWRWKCCKK (Ponnappan et al. 2017) |
| Cyt c(77–101) | GTKMIFVGIKKKEERADLIKKA (Howl and Jones 2015) |
| DAG | cyclic CDAGRKQKC (Mann et al. 2017) |
| D-JNKI-1 | RPKRPTTLNLFPQVPRSQDT (Bonny et al. 2001) |
| DK17 | DRQIKIWFQNRRMKWKK (Bera et al. 2016) |
| DLP | ACKTGSHNQCG (Kumar et al. 2015) |
| DMBT1-derived | GRVEVLYRGSW and GRVRVLYRGSW (Tuttolomondo et al. 2017) |
| dNP2 | KIKKVKKKGRK-KIKKVKKKGRK (Lim et al. 2015) |
| DPV3 | RKKRRRESRKKRRRES (Tacken et al. 2008) |
| DPV1047 | CVKRGLKLRHVRPRVTRMDV (De Coupade et al. 2005) |
| DRTTLTN | DRTTLTN (Gennari et al. 2016) |
| DS4.3 | RIMRILRILKLAR (Jeong et al. 2014) |
| Dynorphin A | YGGFLRRIRPKLKWDNQ (Marinova et al. 2005) |
| EA | GLKKLAELAHKLLKLGC (Yang et al. 2014) |
| EB1 | LIRLWSHLIHIWFQNRRLKWKKK (Lundberg et al. 2007) |
| EF | GLKKLAELFHKLLKLGC (Yang et al. 2014) |
| EHB | RCSHYTGIRCSHMAATTAGIYTGIRCQHVVL-C6H (Cao et al. 2018) |
| EPRNEEK | EPRNEEK (Orihuela et al. 2009) |
| F3** | diphosphorylated dipeptide (Miao et al. 2016) |
| G4R9L4 | G4R9L4 (Ramakrishna et al. 2014) |
| GALA | WEAALAEALAEALAEHLAEALAEALEALAA (Li et al. 2004) |
| GeT | KIAKLKAKIQKLKQKIAKLK (Rakowska et al. 2014) |
| gH625 | HGLASTLTRWAHYNALIRAF (Galdiero et al. 2015) |
| Gi3α(346-355) | KNNLKECGLY (Jones et al. 2005) |
| Glu-Lys | EEEAAKKK (Lewis et al. 2010) |
| GV1001 | EARPALLTSRLRFIPK (Kim et al. 2016a) |
| GWH1 | GYNYAKKLANLAKKFANALW (Serna et al. 2017) |
| H2A derived | SGRGKQGGKARAKAKTRSSRAGLQFPVGRVHRLLRKG (Rosenbluh et al. 2004) |
| H6R6 | H6R6 (Sun et al. 2017) |
| H16 | H16 (Iwasaki et al. 2015) |
| HA2(1-23) | GLFGAIAGFIENGWEGMIDGWYG (Esbjörner et al. 2007) |
| HAIYPRH | HAIYPRH (Shteinfer-Kuzmine et al. 2017) |
| hBD3-3 | GKCSTRGRKCCRRKK (Lee et al. 2015b) |
| HBP | GKRKKKGKGLGKKRDPCLRKYK (Luo et al. 2016) |
| hLF | KCFQWQRNMRKVRGPPVSCIKR (Duchardt et al. 2009) |
| Hph-1 | YARVRRRGPRR (Jung et al. 2011) |
| HR9 | CH5-R9-H5C (Liu et al. 2013a) |
| Hst5 | DSHAKRHHGYKRKFHEKHHSHRGY (Luque-Ortega et al. 2008) |
| I1WL5W | WKKIWSKIKKLLK (Bi et al. 2014) |
| I4WL5W | IKKWWSKIKKLLK (Bi et al. 2014) |
| ID No.2 | MAAWMRSLFSPLKKLWIRMH (Eudes and Macmillan 2014) |
| IMT-P8 | RRWRRWNRFNRRRCR (Gautam et al. 2016) |
| INF | GLFEAIEGFIENGWEGMIDGWYGC (Pichon et al. 1997) |
| iNGR | CRNGRGPDC (Alberici et al. 2013) |
| isl-1 | RVIRVWFQNKRCKDKK (Kilk et al. 2001) |
| JB9 | cskc (Basu and Wickstrom, 1997) |
| JB434 | R9GGLAA-Aib-SGWKH6 (Sangtani et al. 2018) |
| KAFAK | KAFAKLAARLYRKALARQLGVAA (Bartlett et al. 2013) |
| KALA | WEAKLAKALAKALAKHLAKALAKALKACEA (Wyman et al. 1997) |
| Kalata B1 | polycyclic CGETCVGGTCNTPGCTCSWPVCTRNGLPV (Daly et al. 1999) |
| (KFF)3K | (KFF)3K (Rownicki et al. 2017) |
| K-FGF | AAVLLPVLLAAP (Lin et al. 1995) |
| KH | (KH)9 (Chuah et al. 2016) |
| KLA | KLAKLAKKLAKLAK (Huang et al. 2017) |
| KLAK | KLALKLALKALKAALKLA (Oehlke et al. 1998) |
| KLA-R7 | KLAKLAKKLAKLAKGGRRRRRRR (Lemeshko, 2013) |
| KP | MAPTKRKGSCPGAAPNKKP (Villa-Cedillo et al. 2017) |
| KST peptide | STGKANKITITNDKGRLSK (Adachi et al. 2017) |
| L1−6 | PLILLRLLR (Schmidt et al. 2017) |
| L5a | RRWQW (Liu et al. 2016a) |
| L17E | IWLTALKFLGKHAAKHEAKQQLSKL (Akishiba et al. 2017) |
| lactoferrampin(265- 284) | DLIWKLLSKAQEKFGKNKSR (Reyes-Cortes et al. 2017) |
| lactoferricin(17-30) | FKCRRWQWRMKKLG (Reyes-Cortes et al. 2017) |
| lactoferrin(19-40) | KCFMWQEMLNKAGVPKLRCARK (Duchardt et al. 2009) |
| LAH1 | KKLALALALALHALALALALKKA (Moulay et al. 2017) |
| LALF(31-52) | HYRIKPTFRRLKWKYKGKFW (Yanez et al. 2017) |
| LB | FKCRRWQWRMKKLGAPSITCVRRAF) (Liu et al. 2013b) |
| L-CPP | LAGRRRRRRRRRK (Liu et al. 2006) |
| LDP-NLS | KWRRKLKKLRPKKKRKV (Ponnappan and Chugh, 2017) |
| LE10 | LELELELELELELELELELE (Antunes et al. 2013) |
| LF chimera | FKCRRWQWRMKKLG-K-RSKNKGFKEQAKSLLKWILD (Reyes-Cortes et al. 2017) |
| linTT1 | AKRGARSTA (Hunt et al. 2017) |
| LK | LKKLLKLLKKLLKLAG (Kim et al. 2016b) |
| LL37 | LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES (Kim et al. 2016b) |
| LLIIL | LLIIL (Alaybeyoglu et al. 2017) |
| LMWP | VSRRRRRRGGRRRR (Chen et al. 2017d) |
| LP-12 | HIITDPNMAEYL (Kumar et al. 2015) |
| LPAs | RCnRCnK (Gupta et al. 2011) |
| LTV | LTVSPWY (Chopra 2012) |
| lycosin-I | RKGWFKAMKSIAKFIAKEKLKEHL (Tan et al. 2017) |
| Lyp1 | CGNKRTRGC (Fogal et al. 2008) |
| M918 | MVTVLFRRLRIRRACGPPRVRV (El-Andaloussi et al. 2007) |
| Maurocalcine | GDCLPHLKLCKENKGCCSKKCKRRGTNIEKRCR (Poillot et al. 2010) |
| MAP | KLALKLALKALKAALKLA (Oehlke et al. 1998) |
| MAP12 | LKTLTETLKELTKTLTEL (Oehlke et al. 2002) |
| MCoTI-I | polycyclic SGSDGGVCPKILQRCRRDSDCPGACICRGNGYCG (Camarero, 2017) |
| MCoTI-II | polycyclic CPKILKKCRRDSDCPGACICRGNGYCGSGSDGGV (Huang et al. 2015) |
| MFK | MFKLRAKIKVRLRAKIKL (Samuels et al. 2017) |
| Mgpe9 | CRRLRHLRHHYRRRWHRFRC (Vij et al. 2016a) |
| MitP | INLKKLAKL(Aib)KKIL (Howl et al. 2018) |
| m(KLA)-iRGD | klaklakklakla-K-GG-iRGD (Qifan et al. 2016) |
| MMGP1 | MLWSASMRIFASAFSTRGLGTRMLMYCSLPSRCWRK (Pushpanathan et al. 2013) |
| MPER fragment | ELDKWASLWNWFDITNWLWYIK (Song et al. 2009) |
| MPG | GALFLGFLGAAGSTMGA cysteamide (Morris et al. 1997) |
| MPG | GALFLGFLGAAGSTMGASQPKKKRKV cycteamide (Deshayes et al. 2005) |
| MPG-8 | AFLGWLGAWGTMGWSPKKKRK (Crombez et al. 2009b) |
| mRVG | YTIWMPENPRPGTPCDIFTKSRGKRASNGGGRRRRRRRRR (Villa-Cedillo et al. 2017) |
| MT23 | LPKQKRRQRRRM (Zhou et al. 2017) |
| mtCPP1 | r-Dmt-OF (Cerrato et al. 2015) |
| MTM | AAVALLPAVLLALLAP (Fletcher et al. 2010) |
| MTD84 | AVALVAVVAVA (Lim et al. 2014) |
| MTP | MLSLRQSIRFFK (Chuah et al. 2015a, b) |
| MTS | KGEGAAVLLPVLLAAPG (Zhao et al. 2001) |
| MTS1 | AAVLLPVLLAAP (Rojas et al. 1998) |
| Mut3DPT-C9h | VKKKKIKAEIKIYVETLDDIFEQWAHSEDL (de la Torre et al. 2017) |
| Myr-ApoE | Myr-LRKLRKRLLR (Tajik-Ahmadabad et al. 2017) |
| New modalities | Polycyclic, hairpin, stapled peptides for delivery (Valeur et al. 2017, Waldmann et al. 2017) |
| NF1 | Stearyl-AGY(PO3)LLGKTNLKALAALAKKIL (Arukuusk et al. 2013) |
| NF51 | δ-(Stearyl-AGYLLG)OINLKALAALAKKIL (Arukuusk et al. 2013) |
| NF55 | δ-(Stearyl-AGYLLG)OINLKALAALAKAIL (Freimann et al. 2016) |
| NLS | PKKKRKV (Yoneda et al. 1992). |
| NLS-StAx-h | stapled RRWPRXILDXHVRRVWR (Dietrich et al. 2017) |
| NoLS | KKRTLRKNDRKKRC (Yao et al. 2015) |
| Novicidin | KNLRRIIRKGIHIIKKYF (Milosavljevic et al. 2016) |
| NPFSD | VLTNENPFSDP (Gong et al. 2016) |
| NYAD-1 | stapled ITFEDLLDYYGP (Zhang et al. 2008) |
| Oct4-PTD | DVVRVWFCNRRQKGKR (Adachi et al. 2017) |
| P007 | Ac-(RAhxR)4-Ahx-βAla (Greer et al. 2014) |
| P1 | LRRWSLG (Peng et al. 2017b) |
| P2 | WKRTLRRL (Peng et al. 2017b) |
| P3 | YGRKKRRQR (Tan et al. 2006) |
| P7 | RRMKWKK (Watson et al. 2017) |
| P11 | YGRKKRRQRRR (Zhao et al. 2011) |
| P11 | HSDVHK (Bang et al. 2011) |
| P11LRR | P11LRR (Li et al. 2010) |
| P14LRR | (PLPRPR)4 (Brezden et al. 2016) |
| p18 | LSTAADMQGVVTDGMASG (Taylor et al. 2009) |
| P21 | KRKKKGKGLGKKRDPCLRKYK (Dixon et al. 2016) |
| P28 | LSTAADMQGVVTDGMASGLDKDYLKPDD, Leu50-Asp77 of azurin (Yamada et al. 2016) |
| p28 | FLHSGTAVTCTYPALTPQWEGSDCTHRL (Signorelli et al. 2017) |
| p53 peptide MO6 | Stapled TSF*EYWYLL* (Chee et al. 2014) |
| PAF26 | Ac-rkkwfw (Lopez-Garcia et al. 2002) |
| PAS | GKPILFF (Woldetsadik et al. 2017) |
| pCLIP6 | KVRVRVRVpP(pT)RVRERVK (Chen et al. 2017b) |
| pD-SP5 | riPRPSPKMGV(pS)VS (Chen et al. 2017b) |
| PenetraMax | KWFKIQMQIRRWKNKR, L- and D- (Khafagy el et al. 2015) |
| Penetratin | RQIKIWFQNRRMKWKK (Derossi et al. 1994) |
| Pep-1 | KETWWETWWTEWSQPKKKRKV cysteamide (Morris et al. 1997) |
| pepM | KLFMALVAFLRFLTIPPTAGILKRWGTI (Freire et al. 2014) |
| pepR | LKRWGTIKKSKAINVLRGFRKEIGRMLNILNRRRR (Freire et al. 2014) |
| Pept1 | PLILLRLLRGQF (Marks et al. 2011) |
| Peptide 599 | GLFEAIEGFIENGWEGMIDGWYGGGGRRRRRRRRRK (Alexander-Bryant et al. 2015) |
| Pep42 | Cyclic CTVALPGGYVRVC (Kim et al. 2006) |
| PepNeg | SGTQEEY (Neves-Coelho et al. 2017) |
| PepFect6 | Stearyl-AGYLLGK(εTMQ)INLKALAALAKKIL, PF6 (El-Andaloussi et al. 2011) |
| PepFect14 | Stearyl- AGYLLGKLLOOLAAAALOOLL (Ezzat et al. 2011) |
| PG1 | RGGRLCYCRRRFCVCVGR (Liu et al. 2013b) |
| pHLIP | AEQNPIY-WARYADWLFTTPLLLLDLALLV-DADEGT (Andreev et al. 2010) |
| PHPs | H6-H10 peptides (Kimura et al. 2017) |
| PIP1 | RXRRXRRXRIKILFQNRRMKWKK (Ivanova et al. 2008) |
| Pip5e | RXRRBRRXRILFQYRXRBRXRB (Betts et al. 2012) |
| Pip6a | Ac-RXRRBRRXRYQFLIRXRBRXRB (Lehto et al. 2014) |
| POD | CGGG(ARKKAAKA)4 (Dasari et al. 2017) |
| PR9 | FFLIPKG-R9 (Liu et al. 2013a) |
| PTD | YARVRRRGPRRR (Dong et al. 2016) |
| PTD3 | R9-ETWWETWWTEW (Kizaka-Kondoh et al. 2009) |
| PTD4 | YARAAARQARA (McCusker et al. 2007) |
| Poly-Arg | Most popular R7 – R12 (Mitchell et al. 2000, Futaki, 2006) |
| pVEC | LLIILRRRIRKQAHAHSK (Elmquist et al. 2001) |
| Pyrrhocoricin | VDKGSYLPRPTPPRPIYNRN (Otvos et al. 2000) |
| R4K1 | Stapled Ac-RRRRKS*LHRS*LQDS (Speltz et al. 2018) |
| R6dGR | R6dGR (Wang et al. 2017) |
| R8 | R8 (Wender et al. 2001) |
| R8-dGR | R8dGR (Liu et al. 2016b) |
| R9-H4A2 | Ac-YR9-HAHAHH (Okitsu et al. 2017) |
| R6W3 | R6W3 (Bechara et al. 2013) |
| R10W6 | R10W6 (Bechara et al. 2013) |
| RA9 | RRAARRARR (Alhakamy et al. 2013) |
| RALA | WEARLARALARALARHLARALARALRACEA (McCarthy et al. 2014) |
| RDP | CKSVRTWNEI IPSKGCLRVG GRCHPHVNGG GRRRRRRRRC (Xiao et al. 2017) |
| REDV | REDV (Yang et al. 2016) |
| RF | GLKKLARLFHKLLKLGC (Yang et al. 2014) |
| cRGDfC | Cyclic RGDfC (Wada et al. 2017) |
| iRGD | Cyclic CRGDKGPDC (Peng and Kopecek, 2015) |
| RGE | RGERPPR (Yu et al. 2017) |
| RH9 | RRHHRRHRR (Alhakamy et al. 2013) |
| RL9 | RRLLRRLRR (Alhakamy et al. 2013) |
| RL16 | RRLRRLLRRLLRRLRR (Joanne et al. 2009) |
| RT53 | RQIKIWFQNRRMKWKKAKLNAEKLKDFKIRLQYFARGLQV YIRQLRLALQGKT (Jagot-Lacoussiere et al. 2016) |
| RTP004 | RKKRRQRRRG-K15-GRKKRRQRRR) (Lee et al. 2015a) |
| RV24 | RRRRRRRRRGPGVTWTPQAWFQWV (Lo and Wang, 2012) |
| RVG | YTIWMPENPRPGTPCDIFTNSRGKRASNG (Kumar et al. 2007) |
| RVG-9R | YTIWMPENPRPGTPCDIFTNSRGKRASNGGGGRRRRRRRRR (Rassu et al. 2017) |
| RVG29 | YTIWMPENPRPGTPCDIFTNSRGKRASNGGGGRRRRRRRRR (Villa-Cedillo et al. 2017) |
| RW9 | RRWWRRWRR (Alhakamy et al. 2013) |
| RW16 | RRWRRWWRRWWRRWRR (Jobin et al. 2013) |
| (RXR)4 | (R-Ahx-R)4 (Saleh et al. 2010) |
| (rXr)4 | (r-Ahx-r)4 (Vij et al. 2016b) |
| S155 | VKKKKIKREI-KIAAQRYGRELRRMADEFHV (Haidar et al. 2017) |
| S4(13)-PV | ALWKTLLKKVLKAPKKKRKV (Mano et al. 2007) |
| SAP | VRLPPPVRLPPPVRLPPP (Pujals et al. 2006) |
| SAP(E) | VELPPPVELPPPVELPPP (Martin et al. 2011) |
| all-D-SAP | (vrlppp)3 (Pujals et al. 2007) |
| SAPSp-lipo | stearyl-GGGGHGAHEHAGHEHAAGEHHAHE (Suzuki et al. 2017) |
| SAR6EW | SAR6EW (Im et al. 2017) |
| sC18 | GLRKRLRKFRNKIKEK (Oren et al. 1999) |
| (sC18)2 | (GLRKRLRKFRNKIKEK)2 (Gronewold et al. 2017) |
| SMTP motif, | LRLLR (Fuselier and Wimley, 2017) |
| SPACE | Cyclic ACTGSTQHQCG (Hsu and Mitragotri, 2011) |
| SRCRP2-11 | GRVEVLYRGSW (Tuttolomondo et al. 2017) |
| STR-KV | H3K3V6 (Pan et al. 2016) |
| SS-02 | Dmt-r-FK (Alta et al. 2017) |
| SS-20 | F-r-FK (Alta et al. 2017) |
| SS-31 | r-Dmt-KF (Zhao et al. 2005) |
| SynB1 | RGGRLSYSRRRFSTSTGR (Rousselle et al. 2000) |
| T2 | LVGVFH (Kumar et al. 2012) |
| Tat(49-57) | RKKRRQRRR (Vives et al. 1997a) |
| Tat(48-60) | GRKKRRQRRRPPQ (Vives et al. 1997b) |
| Tat(44-57) | CGISYGRKKRRQRRR (Niesner et al. 2002) |
| Tat(37-72) | CFITKALGISYGRKKRRQRRRPPQGSQT-HQVSLSKQ (Fawell et al. 1994) |
| Tat analog | GRKKRRQR (Nguyen et al. 2008) |
| Tat-LK15 | Tat-KLLKLLLKLLLKLLK (Peng et al. 2017a) |
| TCTP | MIIFRALISHKK (Bae et al. 2016) |
| TD-1 | ACSSSPSKHCG (Chen et al. 2006) |
| TD2.2 | SYWYRIVLSRTGRNGRLRVGRERPVLGESP (Heffernan et al. 2012) |
| TH peptide | GYLLGHINLHHLAHL-Aib-HHIL (Chen et al. 2017a) |
| TM2 | PKKGSKKAVTKAQKKDGA (Kochurani et al. 2015) |
| Transportan | GWTLNSAGYLLGKINLKALAALAKKIL, TP (Pooga et al. 1998) |
| TP10 | AGYLLGKINLKALAALAKKIL (Soomets et al. 2000) |
| TPk | VRRFkWWWkFLRR (Bahnsen et al. 2015) |
| Tpl | KWCFRVCYRGICYRRCRGK (Jain et al. 2015) |
| TPP | TKDNNLLGRFELSG (Gehrmann et al. 2014) |
| TT1 | CKRGARSTA (Paasonen et al. 2016) |
| vAMP 059 | INWKKWWQVFYTVV (Dias et al. 2017) |
| vCPP 0769 | RRLTLRQLLGLGSRRRRRSR (Dias et al. 2017) |
| vCPP 2319 | WRRRYRRWRRRRRWRRRPRR (Dias et al. 2017) |
| VDAC(1-26) | MAVPPTYADLGKSARDVFTKGYGFGL (Smilansky et al. 2015) |
| VP22 | NAATATRGRSAASRPTQRPRAPARSASRPRRPVQ (Elliott and O’Hare, 1997) |
| V peptide | TVDNPASTTNKDKLFAVRK (Manosroi et al. 2014) |
| VT5 | DPKGDPKGVTVTVTVTVTGKGDPKPD (Oehlke et al. 1997) |
| W(RW)4 | W(RW)4 (Nasrolahi Shirazi et al. 2013) |
| Xentry | LCLR (Montrose et al. 2014) |
| X-pep | MAARLC (Adachi et al. 2017) |
| YKA | YKALRISRKLAK (Desai et al. 2014) |
| YTA2 | YTAIAWVKAFIRKLRK (Lindgren et al. 2006) |
| YTA4 | IAWVKAFIRKLRKGPLG (Lindgren et al. 2006) |
| Z2 | FWIGGFIKKLKRSKLA (Chen et al. 2017c) |
| Z3 | FKIKKFIGGLWRSKLA (Chen et al. 2017c) |
| Z12 | KRYKNRVASRKCRAKFKQLLQHYREVAAAKSSENDRLRLLLK (Derouazi et al. 2015) |
| ZXR-1 | FKIGGFIKKLWRSKLA (Chen et al. 2017c) |
LifeTein provides the fastest turnaround time and most reliable quality in the industry. Peptides are made in New Jersey, USA. Projects move from conception to bench in only 3–5 days so you can deal with your research deadlines.
Introducing LifeTein‘s faster microwave peptide synthesis technology!
LifeTein’s new platform is designed for maximized speed and efficiency. Unparalleled peptide quality, greater flexibility, and improved reliability make LifeTein the vendor of choice for all your peptide synthesis needs.
Remarkably Fast:
Aβ-(1–42) was dissolved to 1 mM in 100% hexafluoroisopropanol, hexafluoroisopropanol was removed under vacuum, and the peptide was stored at −20 °C. For the aggregation protocols, the peptide was first resuspended in dry Me2SO (DMSO) to 5 mM. For oligomeric conditions, F-12 (without phenol red) culture media was added to bring the peptide to a final concentration of 100 μM, and the peptide was incubated at 4 °C for 24 h. For fibrillar conditions, 10 mM HCl was added to bring the peptide to a final concentration of 100 μM, and the peptide was incubated for 24 h at 37 °C.
ADDLS, amyloid-derived diffusible ligands.
Preparing human islet amyloid polypeptide (hIAPP), also known as amylin, can be challenging due to its hydrophobic amino acid residues.
Here’s an improved method for dissolving lyophilized hIAPP:
To initiate the formation of hIAPP fibrils, introduce the stock solution into the reaction buffer. Conditions for fibril formation were optimized under two pH conditions:
Incubate these samples at 25 °C for several hours.
Reference: JOURNAL OF BIOLOGICAL CHEMISTRY 23965, JULY 8, 2011 VOLUME 286 NUMBER 27
Aducanumab is a human monoclonal antibody that has been studied for the treatment of Alzheimer’s disease.
The Nobel Prize in Physiology or Medicine 2019 was awarded jointly to William G. Kaelin Jr., Sir Peter J. Ratcliffe, and Gregg L. Semenza. LifeTein made one of the key peptides: HIF1a-P564: DLDLEMLAPYIPMDDDFQLR.
A click chemistry was reported about the formation of azides from primary amines. This powerful tool enables the reaction of just one equivalent of a simple diazotizing species, and fluorosulfuryl azide (FSO2N3), for the preparation of over 1,200 azides on 96-well plates in a safe and practical manner. This method greatly expands the number of accessible azides and 1,2,3-triazoles because the primary amine is one of the most abundant functional groups in small compounds, proteins and antibodies.
The method opens the door for numerous applications in drug screening and discovery. The cell penetration peptides can be easily introduced to conjugate with any azide containing drugs, compounds, antibodies, or proteins.
The cell penetration peptides (CPPs) are capable of delivering biologically active cargo to the cell interior. The desired therapeutic cargo could be attached to a CPP using the copper free click chemistry and then delivered to an intracellular target, thereby overcoming the entry restrictions set by the plasma membrane.
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BRAF is an RAF kinase. It is a core component of the RAS/RAF/MEK/ERK signaling cascade, known as mitogen-activated protein kinase (MAPK) pathway. It is one of the major effectors of oncogene RAS, and is often mutated in human cancer cells.
Two FDA approved drugs, Dabrafenib, and vemurafenib, effectively inhibit the most common BRAF variant V600E, a monomeric BRAF. But, the non-V600E BRAF mutations are intrinsically resistant to these drugs. These drugs may also paradoxically stimulate the pathway when the tumor cells contain wild-type BRAF and oncogenic RAS, causing secondary malignancies.
The researchers tried to tackle the dimeric BRAF. The dimeric BRAF, such as the wild type and G469A, a most prevalent non-V600E variant in lung cancer cells, hinges on dimer interface (DIF), a 20aa span near the tail end of the alpha-C helix of BRAF. The researchers designed Braftide using computational modeling, aiming to block the dimerization. They tested the functionality in vitro, in HEK263 cells and colon cancer cell lines.
LifeTein synthesized Braftide (TRHVNILLFM), Null-Braftide (THHVNILLFM), Cy3-Braftide (TRHVNILLFM-Cy3), TAT-Braftide (GRKKRRQRRRPQ-PEG-TRHVNILLFM), and TAT (GRKKRRQRRRPQ). We reviewed here some of the assays that helped support Braftide as an allosteric inhibitor of BRAF dimer and down-regulator of MAPK signaling pathway for cancer therapy.
1) Cell-free in vitro assay: dose-response curve. First of all, the researchers show that Braftide has a sub-micromolar IC50 for dimeric BRAF. Full-length dimeric BRAF-WT and BRAF-G469A (from HEK293F cells) were used for dose-response curves, and the BRAF activity was probed by pMEK production.
2) Cell-free in vitro assay: Saturation binding assay. The researchers used Cy3-labeled Braftide (Cy3-Braftide) to characterize (KD) the binding of Braftide with dimeric BRAF-WT using fluorescence quantification.
3) Cell-free in vitro assay: Immunoprecipitation (IP). The purpose of IP was to show Braftide disrupted the BRAF dimerization. Braftide was added to HEK293 cell lysate coexpressing V5- and FLAG-tagged BRAF-WT. FLAG-tagged BRAF was pulled down by FLAG antibody-conjugated resin, which was further probed for V5-tagged BRAF. Braftide indeed reduced homodimer BRAF.
4) Delivery of Braftide into HEK cell for BRAF inhibition. Braftide was tagged with cell-penetrating peptide TAT. TAT-Braftide (and its negative control TAT alone) was used to treat HEK293 cells transiently transfected with BRAF-WT and BRAF-G469A. Four hours of treatment resulted in reductions of BRAF, pMEK, MEK (i.e. the MAPK pathway), which were analyzed with respective antibodies by immunoblotting.
5) Delivery of Braftide into cancer cells for BRAF inhibition and cell proliferation inhibition. Two colon cancer cell lines (KRAS-G13D-colon carcinoma) were treated with cell-penetrating TAT-Braftide and assayed for the inhibition of BRAF activity, down-regulation of MAPK signaling, and cell proliferation. All were shown positive, while the negative control TAT alone were negative.
https://pubs.acs.org/doi/abs/10.1021/acschembio.9b00191
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Smaller Ions Stabilize β-sheets
