Sets were fit to a simple 1:1 (Langmuir) binding model including a
Sets were fit to a simple 1:1 (Langmuir) binding model including a term for mass transport. The data selection of the binding and dissociation curves for fitting was based on the instruction manual of the BIAcore 3000 instrument, in which 3? s of data at association and dissociation start were eliminated.Additional fileAdditional file 1: Figure S1. Cell penetration of (a) NYAD-41, NYAD-36, NYAD-66 and (b) NYAD-67 and NYAD-1 in 293T cells. Figure S2. Isothermal titration calorimetric (ITC) analyses of the interaction BMS-214662 biological activity betweenCTDM184A/ W185A and NYAD-D36, NYAD-66, NYAD-67 or NYAD-1. Figure S3. i+7 stapled peptides have no effect on HIV-1 release, but impair Gag processing. Figure S4. The Env mutant V120Q/A327P is not resistance to NYAD-1. Figure S5. Kinetic analysis of the interaction between gp120 and (a) NYAD-36, (b) NYAD-66 or (c) NYAD-67 by SPR. Figure S6. Isothermal titration calorimetric (ITC) analyses of the interaction between fulllength cyclic V3 loop peptide and NYAD-41, NYAD-36, NYAD-66, NYAD67 or NYAD-1. Figure S7. Isothermal titration calorimetric (ITC) analyses of the interaction between 15-mer V3 tip peptide and NYAD-36, NYAD-66 or NYAD-67.Competing interests The authors declare that they have no competing interests.Authors’ contributions AKD, EOF and MFS designed experiments, analyzed and prepared the manuscript. HZ designed, performed in vitro assembly assay and biophysical studies by SPR and ITC. AC advised on analysis and interpretation of biophysical study data. FC performed antiviral assays and analyzed the data. AAW designed, performed and analyzed the infectivity and selection experiments. PYM, MM, PB performed the NMR and the in vitro assembly experiments and analyzed the data. DS assisted in antiviral screening. XT and SL prepared all stapled peptides. All authors read and approved the final manuscript.The binding kinetics and affinity of stapled peptides to HIV-1 Yu2gp120 (kindly provided by Dr. Peter Kwong, Vaccine Research Center, NIH) were analyzed by SPR (BIAcore 3000, Piscataway, NJ). The Yu2gp120 or Yu2gp120 protein with V3 loop deletion (Yu2gp120V3) was covalently immobilized to a CM5 sensor chip via amine groups using the amine coupling kit (BIAcore) in 10 mM sodiumAcknowledgments This study was supported by NIH Grant PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/25636517 RO1 AI081604 (AKD) and the intramural fund from the New York Blood Center (AKD). NMR and turbidity studies were supported by NIH Grant R01 AI30917 (MFS). PYM was supported by NIH IMSD grant R25 MBRS-IMSD GM55036 for maximizing doctoral student diversity. This work was supported in part by the Intramural Research Program of the Center for Cancer Research, National Cancer Institute, NIH (EF). We thank Lyudmil Angelov (confocal microscopy) and Yelena Oksov (electron microscopy) for their technical help. We thank K. Waki for constructing the pCMVdeltaR8.2/ PR- clone and. J. Burns for providing the VSV-G expression vector. HIV-Ig was obtained from the NIH AIDS Research and Reference Reagent Program.Zhang et al. Retrovirology 2013, 10:136 http://www.retrovirology.com/content/10/1/Page 19 ofAuthor details 1 Laboratory of Molecular Modeling, Drug PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/28192408 Design, Lindsley F. Kimball Research Institute of the New York Blood Center, 310 E 67th Street, New York, NY 10065, USA. 2Virus-Cell Interaction Section, HIV Drug Resistance Program, National Cancer Institute-Frederick, Frederick, MD 21702, USA. 3Howard Hughes Medical Institute and Department of Chemistry and Biochemistry, University of Maryland, Baltimore.