A systematic proximity ligation approach to studying protein-substrate specificity identifies the substrate spectrum of the Ssh1 translocon

Nir Cohen, Naama Aviram, Maya Schuldiner

نتاج البحث: نشر في مجلةمقالةمراجعة النظراء

ملخص

Many cellular functions are carried out by protein pairs or families, providing robustness alongside functional diversity. For such processes, it remains a challenge to map the degree of specificity versus promiscuity. Protein–protein interactions (PPIs) can be used to inform on these matters as they highlight cellular locals, regulation and, in cases where proteins affect other proteins - substrate range. However, methods to systematically study transient PPIs are underutilized. In this study, we create a novel approach to systematically compare stable or transient PPIs between two yeast proteins. Our approach, Cel-lctiv (CELlular biotin-Ligation for Capturing Transient Interactions in vivo), uses high-throughput pairwise proximity biotin ligation for comparing PPIs systematically and in vivo. As a proof of concept, we studied the homologous translocation pores Sec61 and Ssh1. We show how Cel-lctiv can uncover the unique substrate range for each translocon allowing us to pinpoint a specificity determinator driving interaction preference. More generally, this demonstrates how Cel-lctiv can provide direct information on substrate specificity even for highly homologous proteins.

اللغة الأصليةالإنجليزيّة
رقم المقالe113385
عدد الصفحات14
دوريةEMBO Journal
مستوى الصوت42
رقم الإصدار11
المعرِّفات الرقمية للأشياء
حالة النشرنُشِر - 1 يونيو 2023

All Science Journal Classification (ASJC) codes

  • !!General Immunology and Microbiology
  • !!General Biochemistry, Genetics and Molecular Biology
  • !!Molecular Biology
  • !!General Neuroscience

بصمة

أدرس بدقة موضوعات البحث “A systematic proximity ligation approach to studying protein-substrate specificity identifies the substrate spectrum of the Ssh1 translocon'. فهما يشكلان معًا بصمة فريدة.

قم بذكر هذا