MST and TRIC Technology to Reliably Study PROTAC Binary and Ternary Binding in Drug Development

Tanja Bartoschik; Andreas Zoephel; Klaus Rumpel; Alessio Ciulli; Charles Heffern

doi:10.1007/978-1-0716-1665-9_6

MST and TRIC Technology to Reliably Study PROTAC Binary and Ternary Binding in Drug Development

Tanja Bartoschik, Andreas Zoephel, Klaus Rumpel, Alessio Ciulli, Charles Heffern (Lead / Corresponding author)

Biological Chemistry and Drug Discovery

Research output: Chapter in Book/Report/Conference proceeding › Chapter (peer-reviewed) › peer-review

7 Citations (Scopus)

387 Downloads (Pure)

Abstract

PROTACs have shown promise as a new class of therapy, with a unique mechanism of action orthogonal to traditional small molecules that are used to regulate protein activity. Their novel MOA utilizing the body's natural protein degradation machinery degrades a protein of interest rather than inhibiting its function. This strategy has several advantages over conventional small-molecule inhibitors, e.g., higher sensitivity, less off-target effects, and greater target space. However, unlocking the potential of PROTACs necessitates drug discovery techniques that can support the complexity of the novel MOA. In this chapter, we describe the application of MicroScale Thermophoresis (MST) and Temperature-Related Intensity Change (TRIC) to characterize both the binary and ternary binding of PROTACs with target proteins and ubiquitin ligases along with an efficient determination of the cooperativity of the ternary complex formation. The assay development and experimental procedure to characterize the well-described BET PROTAC MZ1 show how MST and TRIC can be applied as a fast and highly sensitive method for PROTAC discovery.

Original language	English
Title of host publication	Targeted Protein Degradation
Subtitle of host publication	Methods and Protocols
Editors	Angela M. Cacace, Christopher M. Hickey, Miklós Békés
Place of Publication	New York
Publisher	Humana Press
Pages	115-133
Number of pages	19
Edition	1
ISBN (Electronic)	9781071616659
ISBN (Print)	9781071616642 (hbk), 9781071616673 (pbk)
DOIs	https://doi.org/10.1007/978-1-0716-1665-9_6
Publication status	Published - 2021

Publication series

Name	Methods in Molecular Biology
Volume	2365
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Keywords

MST
MicroScale Thermophoresis
PROTAC
TRIC
Temperature-related intensity change

ASJC Scopus subject areas

Molecular Biology
Genetics

Access to Document

10.1007/978-1-0716-1665-9_6

Author Accepted ManuscriptAccepted author manuscript, 1.79 MB

Cite this

Bartoschik, T., Zoephel, A., Rumpel, K., Ciulli, A., & Heffern, C. (2021). MST and TRIC Technology to Reliably Study PROTAC Binary and Ternary Binding in Drug Development. In A. M. Cacace, C. M. Hickey, & M. Békés (Eds.), Targeted Protein Degradation: Methods and Protocols (1 ed., pp. 115-133). (Methods in Molecular Biology; Vol. 2365). Humana Press. https://doi.org/10.1007/978-1-0716-1665-9_6

@inbook{ecb682fdeb93428e94620fe213d91346,

title = "MST and TRIC Technology to Reliably Study PROTAC Binary and Ternary Binding in Drug Development",

abstract = "PROTACs have shown promise as a new class of therapy, with a unique mechanism of action orthogonal to traditional small molecules that are used to regulate protein activity. Their novel MOA utilizing the body's natural protein degradation machinery degrades a protein of interest rather than inhibiting its function. This strategy has several advantages over conventional small-molecule inhibitors, e.g., higher sensitivity, less off-target effects, and greater target space. However, unlocking the potential of PROTACs necessitates drug discovery techniques that can support the complexity of the novel MOA. In this chapter, we describe the application of MicroScale Thermophoresis (MST) and Temperature-Related Intensity Change (TRIC) to characterize both the binary and ternary binding of PROTACs with target proteins and ubiquitin ligases along with an efficient determination of the cooperativity of the ternary complex formation. The assay development and experimental procedure to characterize the well-described BET PROTAC MZ1 show how MST and TRIC can be applied as a fast and highly sensitive method for PROTAC discovery.",

keywords = "MST, MicroScale Thermophoresis, PROTAC, TRIC, Temperature-related intensity change",

author = "Tanja Bartoschik and Andreas Zoephel and Klaus Rumpel and Alessio Ciulli and Charles Heffern",

note = "{\textcopyright} 2021. Springer Science+Business Media, LLC, part of Springer Nature.",

year = "2021",

doi = "10.1007/978-1-0716-1665-9_6",

language = "English",

isbn = "9781071616642 (hbk)",

series = "Methods in Molecular Biology",

publisher = "Humana Press",

pages = "115--133",

editor = "Cacace, {Angela M.} and Hickey, {Christopher M.} and Mikl{\'o}s B{\'e}k{\'e}s",

booktitle = "Targeted Protein Degradation",

address = "United States",

edition = "1",

}

Bartoschik, T, Zoephel, A, Rumpel, K, Ciulli, A & Heffern, C 2021, MST and TRIC Technology to Reliably Study PROTAC Binary and Ternary Binding in Drug Development. in AM Cacace, CM Hickey & M Békés (eds), Targeted Protein Degradation: Methods and Protocols. 1 edn, Methods in Molecular Biology, vol. 2365, Humana Press, New York, pp. 115-133. https://doi.org/10.1007/978-1-0716-1665-9_6

MST and TRIC Technology to Reliably Study PROTAC Binary and Ternary Binding in Drug Development. / Bartoschik, Tanja; Zoephel, Andreas; Rumpel, Klaus et al.
Targeted Protein Degradation: Methods and Protocols. ed. / Angela M. Cacace; Christopher M. Hickey; Miklós Békés. 1. ed. New York: Humana Press, 2021. p. 115-133 (Methods in Molecular Biology; Vol. 2365).

Research output: Chapter in Book/Report/Conference proceeding › Chapter (peer-reviewed) › peer-review

TY - CHAP

T1 - MST and TRIC Technology to Reliably Study PROTAC Binary and Ternary Binding in Drug Development

AU - Bartoschik, Tanja

AU - Zoephel, Andreas

AU - Rumpel, Klaus

AU - Ciulli, Alessio

AU - Heffern, Charles

PY - 2021

Y1 - 2021

N2 - PROTACs have shown promise as a new class of therapy, with a unique mechanism of action orthogonal to traditional small molecules that are used to regulate protein activity. Their novel MOA utilizing the body's natural protein degradation machinery degrades a protein of interest rather than inhibiting its function. This strategy has several advantages over conventional small-molecule inhibitors, e.g., higher sensitivity, less off-target effects, and greater target space. However, unlocking the potential of PROTACs necessitates drug discovery techniques that can support the complexity of the novel MOA. In this chapter, we describe the application of MicroScale Thermophoresis (MST) and Temperature-Related Intensity Change (TRIC) to characterize both the binary and ternary binding of PROTACs with target proteins and ubiquitin ligases along with an efficient determination of the cooperativity of the ternary complex formation. The assay development and experimental procedure to characterize the well-described BET PROTAC MZ1 show how MST and TRIC can be applied as a fast and highly sensitive method for PROTAC discovery.

AB - PROTACs have shown promise as a new class of therapy, with a unique mechanism of action orthogonal to traditional small molecules that are used to regulate protein activity. Their novel MOA utilizing the body's natural protein degradation machinery degrades a protein of interest rather than inhibiting its function. This strategy has several advantages over conventional small-molecule inhibitors, e.g., higher sensitivity, less off-target effects, and greater target space. However, unlocking the potential of PROTACs necessitates drug discovery techniques that can support the complexity of the novel MOA. In this chapter, we describe the application of MicroScale Thermophoresis (MST) and Temperature-Related Intensity Change (TRIC) to characterize both the binary and ternary binding of PROTACs with target proteins and ubiquitin ligases along with an efficient determination of the cooperativity of the ternary complex formation. The assay development and experimental procedure to characterize the well-described BET PROTAC MZ1 show how MST and TRIC can be applied as a fast and highly sensitive method for PROTAC discovery.

KW - MST

KW - MicroScale Thermophoresis

KW - PROTAC

KW - TRIC

KW - Temperature-related intensity change

UR - http://www.scopus.com/inward/record.url?scp=85114089102&partnerID=8YFLogxK

U2 - 10.1007/978-1-0716-1665-9_6

DO - 10.1007/978-1-0716-1665-9_6

M3 - Chapter (peer-reviewed)

C2 - 34432241

SN - 9781071616642 (hbk)

SN - 9781071616673 (pbk)

T3 - Methods in Molecular Biology

SP - 115

EP - 133

BT - Targeted Protein Degradation

A2 - Cacace, Angela M.

A2 - Hickey, Christopher M.

A2 - Békés, Miklós

PB - Humana Press

CY - New York

ER -

Bartoschik T, Zoephel A, Rumpel K, Ciulli A, Heffern C. MST and TRIC Technology to Reliably Study PROTAC Binary and Ternary Binding in Drug Development. In Cacace AM, Hickey CM, Békés M, editors, Targeted Protein Degradation: Methods and Protocols. 1 ed. New York: Humana Press. 2021. p. 115-133. (Methods in Molecular Biology). Epub 2021 Aug 26. doi: 10.1007/978-1-0716-1665-9_6

MST and TRIC Technology to Reliably Study PROTAC Binary and Ternary Binding in Drug Development

Abstract

Publication series

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this