Engineering Antibodies and Alternative Binders for Therapeutic Uses

Aoki, Wataru

doi:10.1007/978-981-13-5868-5_10

Cited by 2 publications

(2 citation statements)

References 169 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In recent years, yeast surface display has demonstrated immense versatility as a platform for protein engineering 26–28 . Among many applications, the yeast endoplasmic reticulum sequestration screening (YESS) system provides the ability to probe enzyme–substrate interactions 29–31 .…”

Section: Introductionmentioning

confidence: 99%

“…In recent years, yeast surface display has demonstrated immense versatility as a platform for protein engineering. [26][27][28] Among many applications, the yeast endoplasmic reticulum sequestration screening (YESS) system provides the ability to probe enzyme-substrate interactions. [29][30][31] In this system, an enzyme and a substrate fused to the C-terminus of Aga2p are co-localized in the yeast endoplasmic reticulum (ER) by ER targeting signal sequences and retained by fusion to peptides capable of binding KDEL receptors.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Endoplasmic reticulum sequestration empowers phosphorylation profiling on the yeast surface

et al. 2022

View full text Add to dashboard Cite

Cells sense cues in their surrounding environment, turning extracellular stimuli into intracellular responses. Transient posttranslational modifications, such as phosphorylation, often guide these responses through tightly regulated signaling networks.There is great interest in understanding phosphorylation events that yield responses in cell signaling, but to date, methods to characterize the phosphoproteome rely on low-throughput methods. To overcome this limitation, this study optimizes a yeast surface display system for high-throughput screening of kinase-substrate interaction using endoplasmic reticulum (ER) sequestration. We demonstrate the ability of yeast to display phosphorylated full-length receptor cytoplasmic domains and additionally optimized magnetic bead selections to efficiently recover yeast displaying phosphorylated domains. These lessons were applied to profiling the substrate specificity of the tyrosine kinase lymphocyte cell-specific protein-tyrosine kinase by mutating a phosphoacceptor motif within the CD28 cytoplasmic domain and to building multienzyme phosphorylation cascades in the yeast ER. Collectively, this study provides a facile approach for profiling kinase-substrate reactions in high throughput.

show abstract

Section: Introductionmentioning

confidence: 99%