JoVE Journal

Immunology and Infection

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Peptide-based Identification of Functional Motifs and their Binding Partners


12.3K Views

14:28 min

June 30th, 2013

June 30th, 2013

12,315 Views

0:05

Title

2:55

Nef Scanning Peptide TUNEL Assay

4:43

Competitive Inhibition Analysis Using Peptides

6:02

Isolating Motif Binding Partners Using Peptides as Bait

9:25

Nef-GFP Secretion Fluorescent Plate Reader Assay of Antibody Inhibition

10:33

Identification of Nef Functional Motifs and their Binding Partners

13:23

Conclusion

Techniques to dissect the mechanisms underlying the secretion of HIV-1 Nef in exosomes are described. Specific short peptides derived from Nef and protein transfection were exploited to determine the structure, function, and binding partners of Nef’s Secretion Modification Region. These procedures have general relevance in many mechanistic studies.

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