TY - JOUR
T1 - Engineering Selective TIMPs Using a Counter-Selective Screening Strategy
AU - Ahmadighadykolaei, Hannaneh
AU - Radisky, Evette S.
AU - Raeeszadeh-Sarmazdeh, Maryam
N1 - Publisher Copyright:
© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2024
Y1 - 2024
N2 - The yeast surface display platform provides a powerful approach for screening protein diversity libraries to identify binders with an enhanced affinity toward a binding partner. Here, we describe an adaptation of the approach to identify binders with enhanced specificity toward one among multiple closely related binding partners. Specifically, we describe methods for engineering selective matrix metalloproteinase (MMP) inhibitors via yeast surface display of a tissue inhibitor of metalloproteinase (TIMP) diversity library coupled with a counter-selective screening strategy. This protocol may also be employed for developing selective protein binders or inhibitors toward other targets.
AB - The yeast surface display platform provides a powerful approach for screening protein diversity libraries to identify binders with an enhanced affinity toward a binding partner. Here, we describe an adaptation of the approach to identify binders with enhanced specificity toward one among multiple closely related binding partners. Specifically, we describe methods for engineering selective matrix metalloproteinase (MMP) inhibitors via yeast surface display of a tissue inhibitor of metalloproteinase (TIMP) diversity library coupled with a counter-selective screening strategy. This protocol may also be employed for developing selective protein binders or inhibitors toward other targets.
KW - Directed evolution
KW - Enzyme inhibitors
KW - Extracellular matrix
KW - Fluorescence-activated cell sorting
KW - Matrix metalloproteinase
KW - Rational design of proteins
KW - Tissue inhibitor of metalloproteinases
KW - Yeast surface display
UR - http://www.scopus.com/inward/record.url?scp=85178502935&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85178502935&partnerID=8YFLogxK
U2 - 10.1007/978-1-0716-3589-6_20
DO - 10.1007/978-1-0716-3589-6_20
M3 - Article
C2 - 38038946
AN - SCOPUS:85178502935
SN - 1064-3745
VL - 2747
SP - 257
EP - 278
JO - Methods in molecular biology (Clifton, N.J.)
JF - Methods in molecular biology (Clifton, N.J.)
ER -