"..folded HD5 was generated from a synthesized 80% pure linear peptide (CPC Scientific, Sunnyvale, CA) by thiol disulfide reshuffling overnight at room temperature in the presence of 3 mM reduced and 0.3 mM oxidized glutathione, 2 M guanidine hydrochloride, and 0.25 M sodium bicarbonate, pH 8.3, at a peptide concentration of 0.25 mg/ml [..] All α-defensins were quantified by UV absorbance at 280 nm using calculated molar extinction coefficients (18)."
Abstract
Human α-defensins, such as human α-defensin 5 (HD5), block infection of non-enveloped viruses, including human adenoviruses (AdV), papillomaviruses (HPV), and polyomaviruses. Through mutational analysis of HD5, we have identified arginine residues that contribute to antiviral activity against AdV and HPV. Of two arginine residues paired on one face of HD5, Arg-28 is critical for both viruses, while Arg-9 is only important for AdV. Two arginine residues on the opposite face of the molecule (Arg-13 and Arg-32) and unpaired Arg-25 are less important for both. In addition, hydrophobicity at residue 29 is a major determinant of anti-adenoviral activity, and a chemical modification that prevents HD5 self-association was strongly attenuating. Although HD5 binds to the capsid of AdV, the molecular basis for this interaction is undefined. Capsid binding by HD5 is not purely charge-dependent, as substitution of lysine for Arg-9 and Arg-28 was deleterious. Analysis of HD5 analogs that retained varying levels of potency demonstrated that anti-adenoviral activity is directly correlated with HD5 binding to the virus, confirming that the viral capsid rather than the cell is the relevant target. Also, AdV aggregation induced by HD5 binding is not sufficient for neutralization. Rather, these studies confirm that the major mechanism of HD5-mediated neutralization of AdV depends upon specific binding to the viral capsid through interactions mediated in part by critical arginine residues, hydrophobicity at residue 29, and multimerization of HD5, which increases initial binding of virus to the cell but prevents subsequent viral uncoating and genome delivery to the nucleus.
SOCIAL MEDIA
Connect with us and stay updated by following our social media channels.
Latest Briefings from our Knowledge Center
Press Releases, Industry News, Articles, and Technical Content
Boehm, M., Beaumont, K., Jones, R.M., Kalgutkar, A.S., Zhang, L., Atkinson, K., Bai, G., Brown, J.A., Eng, H., Goetz, G.H. and Holder, B.R. Journal of Medicinal Chemistry (2017).
- Pfizer Worldwide Research & Development, Cambridge, Massachusetts 02139, United States
- Pfizer Worldwide Research & Development, Groton, Connecticut 06340, United States
The chemokine receptor CXCR7 is an attractive target for a variety of diseases. While several small-molecule modulators of CXCR7 have been reported, peptidic macrocycles may provide advantages in terms of potency, selectivity, and reduced off-target activity
Bainbridge, Travis W., et al. Scientific Reports 7.1 (2017): 12524.
- Protein Chemistry, Molecular Oncology, Discovery Chemistry, Cancer Immunology, and Neuroscience. Genentech Inc., South San Francisco, CA, 94080, USA
"ANP FAP and aP NIRF were synthesized by CPC Scientific (Sunnyvale, CA)… Another internally-quenched FRET peptide substrate (ANPFAP) for FAP has recently been reported and demonstrated for use as an activity-based, in vivo imaging tool. The peptide sequence contains two internal Gly-Pro dipeptide motifs, susceptible to FAP cleavage and a Cy5.5/QSY21, quenched-FRET pair."
Coorens, Maarten, et al. The Journal of Immunology 199.4 (2017): 1418-1428.
"CATH-2 and LL-37 were synthesized by Fmoc-chemistry at CPC Scientific (Sunnyvale, CA)."
Tcholakov, I., Grimshaw, C.E., Shi, L., Kiryanov, A., Murphy, S.T., Larson, C.J., Plonowski, A. and Ermolieff, J. Bioscience Reports 37, no. 3 (2017): BSR20170275.
- Departments of In Vitro Pharmacology, Immunology, Enzymology and Biophysical Chemistry, Medicinal Chemistry, External Innovation, Metabolic Disease, In Vitro Pharmacology, and Gastrointestinal and Enterology Discovery Unit, Takeda California, Inc., 10410 Science Center Drive, San Diego, CA 92121, U.S.A
The substrate used for our PHD2 kinetic study was a 17-mer peptide mimicking the sequence of HIF-1a surrounding the Pro564 residue hydroxylated by the PHD enzymes (Biotin-DLEMLAPYIPMDDDFQL). The substrate used for FIH1 was a 34-mer peptide mimicking the sequence of HIF-1α surrounding residue Asn803 (DESGLPQLTSYDCEVNAPIQGSRNLLQGEELLRAL). Both peptides were synthesized by CPC Scientific Inc.
Wilson, Sarah S., et al. PLoS Pathogens 13.6 (2017): e1006446.
"Cryptdin 2 (UniProtKB: P28309, LRDLVCYCRTRGCKRRERMNGTCRKGHLMYTLCCR)... obtained by oxidative refolding of partially purified linear peptides (synthesized by CPC Scientific...) and purifying the correctly folded species by reverse-phase high-pressure liquid chromatography (RP-HPLC). Purity was determined by analytical RP-HPLC, and the mass of the disulfide-bonded peptides was verified by high mass accuracy liquid chromatography-mass spectrometry."
Sattiraju, A., et al. Oncotarget 2017, 8 (26), 42997-43007.
For Ac-225 labeling, the prepared DOTA-Pep-1L (CPC-scientific, San Jose, CA) was incubated with Ac-225 at 70°C for 50 minutes. The TLC plates were scanned on a BioScan Imaging Scanner. Cu-64 was purchased from Washington University in St. Louis. The custom peptide specific to IL13RA2 and a scrambled peptide were conjugated with NOTA by CPC scientific Inc (San Jose, CA). Both the peptides, Pep-1L and scrambled peptide-NOTA were radiolabeled with Cu-64 according to the previously reported methods [15].
Puthenveetil, Sujiet, et al. PloS One 12.5 (2017): e0178452.
- Worldwide Medicinal Chemistry, Pfizer Global R&D, Groton, Connecticut, United States of America.
- Pfizer Oncology Research, Pearl River, NY, United States of America.
"Peptide-payload conjugate (7) was prepared by reacting 2mM aizoacetyl-Ser-Lys-Gly-Ser-Lys (CPC scientific, inc. Sunnywale, CA) with 8 mM NHS-ester payload [19] in 50% Dimethyl sulfoxide (DMSO), 50 mM borate buffer pH 8.5 for 2 h at 37°C."
Kwon, Ester J., et al. Advanced Materials 29.35 (2017).
"[..] synthesized for initial screening with FAM-conjugated lysine at the C-terminal end of the membrane-interactive peptide and with or without D[KLAKLAK]2 on the C-terminus using standard Fmoc chemistry[..] All peptides were synthesized with N-terminal myristic acid and C-terminal amine. [..] larger quantities of the peptides were synthesized by CPC Scientific to 90% purity"
Noland, Cameron L., et al. Proceedings of the National Academy of Sciences 114.30 (2017): E6044-E6053.
- Departments of Structural Biology, Infectious Diseases, Biochemical and Cellular Pharmacology, Pathology, Translational Immunology, and Biomolecular Resource Group, Genentech, Inc., South San Francisco, CA 94080.
"..peptide substrates were synthesized based on the Pal lipoprotein, Pal peptideshort (Pam2Cys-SSNKNGGK-Biotin; CPC Scientific, Inc.).."