Boston, Massachusetts, United States
Contact Info
3K followers
500+ connections
About
Activity
-
This platform may crash harder than #crowdstrike on July 24th 🔥 Tune in to watch Morgan J Ingram, Will Falkenborg 🌭, and yours truly talk about…
This platform may crash harder than #crowdstrike on July 24th 🔥 Tune in to watch Morgan J Ingram, Will Falkenborg 🌭, and yours truly talk about…
Liked by Stefano Gullà
-
Abscopal effect from intratumoral anchored cytokine therapy. https://lnkd.in/ez4CtKzp
Abscopal effect from intratumoral anchored cytokine therapy. https://lnkd.in/ez4CtKzp
Liked by Stefano Gullà
Experience
Education
Publications
-
Designed BH3 Peptides with High Affinity and Specificity for Targeting Mcl-1 in Cells
ACS Chemical Biology
Mcl-1 is overexpressed in many cancers and can confer resistance to cell-death signaling in refractory disease. Molecules that specifically inhibit Mcl-1 hold potential for diagnosing and disrupting Mcl-1-dependent cell survival. We selected three peptides from a yeast-surface display library that showed moderate specificity and affinity for binding to Mcl-1 over Bfl-1, Bcl-xL, Bcl-2, and Bcl-w. Specificity for Mcl-1 was improved by introducing threonine at peptide position 2e. The most…
Mcl-1 is overexpressed in many cancers and can confer resistance to cell-death signaling in refractory disease. Molecules that specifically inhibit Mcl-1 hold potential for diagnosing and disrupting Mcl-1-dependent cell survival. We selected three peptides from a yeast-surface display library that showed moderate specificity and affinity for binding to Mcl-1 over Bfl-1, Bcl-xL, Bcl-2, and Bcl-w. Specificity for Mcl-1 was improved by introducing threonine at peptide position 2e. The most specific peptide, MS1, bound Mcl-1 with 40-fold or greater specificity over four other human Bcl-2 paralogs. In BH3 profiling assays, MS1 caused depolarization in several human Mcl-1-dependent cell lines with EC50 values of 3 μM, contrasted with EC50 values of >100 μM for Bcl-2-, Bcl-xL-, or Bfl-1-dependent cell lines. MS1 is at least 30-fold more potent in this assay than the previously used Mcl-1 targeting reagent NoxaA BH3. These peptides can be used to detect Mcl-1 dependency in cells and provide leads for developing Mcl-1 targeting therapeutics.
Other authorsSee publication -
In silico and in vitro elucidation of BH3 binding specificity toward Bcl-2
Biochemistry
Interactions between Bcl-2 like proteins and BH3 domains play a key role in the regulation of apoptosis. Despite the overall structural similarity of their interaction with helical BH3 domains, Bcl-2 like proteins exhibit an intricate spectrum of binding specificities whose underlying basis is not well understood. Here, we characterize these interactions using Rosetta FlexPepBind, a protocol for the prediction of peptide binding specificity that evaluates the binding potential of different…
Interactions between Bcl-2 like proteins and BH3 domains play a key role in the regulation of apoptosis. Despite the overall structural similarity of their interaction with helical BH3 domains, Bcl-2 like proteins exhibit an intricate spectrum of binding specificities whose underlying basis is not well understood. Here, we characterize these interactions using Rosetta FlexPepBind, a protocol for the prediction of peptide binding specificity that evaluates the binding potential of different peptides based on structural models of the corresponding peptide-receptor complexes. For two prominent players, Bcl-xL and Mcl-1, we obtain good agreement with a large set of experimental SPOT array measurements and recapitulate the binding specificity of peptides derived by yeast display in a previous study. We extend our approach to a third member of this family, Bcl-2: we test our blind prediction of the binding of 180 BIM-derived peptides with a corresponding experimental SPOT array. Both prediction and experiment reveal a Bcl-2 binding specificity pattern that resembles that of Bcl-xL. Finally, we extend this application to accurately predict the specificity pattern of additional human BH3-only derived peptides. This study characterizes the distinct patterns of binding specificity of BH3-only derived peptides for the Bcl-2 like proteins Bcl-xL, Mcl-1 and Bcl-2, and provides insight into the structural basis of determinants of specificity.
-
Synthesis of a spin-labeled anti-estrogen as a dynamic motion probe for the estrogen receptor ligand binding domain
The preparation and characterization of a novel nitroxide spin probe based on a steroidal anti-estrogen is described. The probe 5 demonstrated very high binding affinity for both the alpha and beta isoforms of the estrogen receptor-ligand binding domain. EPR spectrometric studies demonstrate conformational constraints for the ligand, consistent with the nitroxyl moiety occupying a position just beyond the receptor-solvent interface.
-
Determinants of BH3 binding specificity for Mcl-1 versus Bcl-xL
Interactions among Bcl-2 family proteins are important for regulating apoptosis. Prosurvival members of the family interact with proapoptotic BH3 (Bcl-2-homology-3)-only members, inhibiting execution of cell death through the mitochondrial pathway. Structurally, this interaction is mediated by binding of the alpha-helical BH3 region of the proapoptotic proteins to a conserved hydrophobic groove on the prosurvival proteins. Native BH3-only proteins exhibit selectivity in binding prosurvival…
Interactions among Bcl-2 family proteins are important for regulating apoptosis. Prosurvival members of the family interact with proapoptotic BH3 (Bcl-2-homology-3)-only members, inhibiting execution of cell death through the mitochondrial pathway. Structurally, this interaction is mediated by binding of the alpha-helical BH3 region of the proapoptotic proteins to a conserved hydrophobic groove on the prosurvival proteins. Native BH3-only proteins exhibit selectivity in binding prosurvival members, as do small molecules that block these interactions. Understanding the sequence and structural basis of interaction specificity in this family is important, as it may allow the prediction of new Bcl-2 family associations and/or the design of new classes of selective inhibitors to serve as reagents or therapeutics. In this work, we used two complementary techniques--yeast surface display screening from combinatorial peptide libraries and SPOT peptide array analysis--to elucidate specificity determinants for binding to Bcl-x(L)versus Mcl-1, two prominent prosurvival proteins. We screened a randomized library and identified BH3 peptides that bound to either Mcl-1 or Bcl-x(L) selectively or to both with high affinity. The peptides competed with native ligands for binding into the conserved hydrophobic groove, as illustrated in detail by a crystal structure of a specific peptide bound to Mcl-1. Mcl-1-selective peptides from the screen were highly specific for binding Mcl-1 in preference to Bcl-x(L), Bcl-2, Bcl-w, and Bfl-1, whereas Bcl-x(L)-selective peptides showed some cross-interaction with related proteins Bcl-2 and Bcl-w. Mutational analyses using SPOT arrays revealed the effects of 170 point mutations made in the background of a peptide derived from the BH3 region of Bim, and a simple predictive model constructed using these data explained much of the specificity observed in our Mcl-1 versus Bcl-x(L) binders.
-
Mcl-1-Bim complexes accommodate surprising point mutations via minor structural changes
Mcl-1 is an antiapoptotic Bcl-2-family protein that protects cells against death. Structures of Mcl-1, and of other anti-apoptotic Bcl-2 proteins, reveal a surface groove into which the alpha-helical BH3 regions of certain proapoptotic proteins can bind. Despite high overall structural conservation, differences in this groove afford binding specificity that is important for the mechanism of Bcl-2 family function. We report the crystal structure of human Mcl-1 bound to a BH3 peptide derived from…
Mcl-1 is an antiapoptotic Bcl-2-family protein that protects cells against death. Structures of Mcl-1, and of other anti-apoptotic Bcl-2 proteins, reveal a surface groove into which the alpha-helical BH3 regions of certain proapoptotic proteins can bind. Despite high overall structural conservation, differences in this groove afford binding specificity that is important for the mechanism of Bcl-2 family function. We report the crystal structure of human Mcl-1 bound to a BH3 peptide derived from human Bim and the structures for three complexes that accommodate large physicochemical changes at conserved Bim sites. The mutations had surprisingly modest effects on complex stability, and the structures show that Mcl-1 can undergo small changes to accommodate the mutant ligands. For example, a shift in a leucine side chain fills a hole left by an isoleucine-to-alanine mutation at the first hydrophobic buried position of Bim BH3. Larger changes are also observed, with shifting of helix alpha3 accommodating an isoleucine-to-tyrosine mutation at this same position. We surveyed the variation in available Mcl-1 and Bcl-x(L) structures and observed moderate flexibility that is likely critical for facilitating interactions of diverse BH3-only proteins with Mcl-1. With the antiapoptotic Bcl-2 family members attracting significant attention as therapeutic targets, these structures contribute to our growing understanding of how specificity is achieved and can help to guide the design of novel inhibitors that target Mcl-1.
-
Molecular-scale force measurement in a coiled-coil peptide dimer by electron spin resonance
A new method for measuring forces between small protein domains based on double electron-electron resonance (DEER) spectroscopy is demonstrated using a model peptide derived from the alpha-helical coiled-coil leucine zipper of yeast transcriptional activator GCN4. The equilibrium distribution of distances between two nitroxide spin labels rigidly attached to the helices of the dimer was determined by DEER and yielded a closing force of 100 +/- 10 pN between monomers, in excellent agreement with…
A new method for measuring forces between small protein domains based on double electron-electron resonance (DEER) spectroscopy is demonstrated using a model peptide derived from the alpha-helical coiled-coil leucine zipper of yeast transcriptional activator GCN4. The equilibrium distribution of distances between two nitroxide spin labels rigidly attached to the helices of the dimer was determined by DEER and yielded a closing force of 100 +/- 10 pN between monomers, in excellent agreement with theoretical predictions.
Patents
-
KLRG1 DEPLETING ANTIBODIES
Issued US11180561B2
-
CD21 Antibodies and Uses Thereof
Filed WO2023283211A2
Anti-CD21 antibodies for treatment of autoimmune disorders.
-
ANTI-KLRG1 ANTIBODIES
Filed US20210347899A1
Languages
-
Italian
Native or bilingual proficiency
More activity by Stefano
-
🌟 Speaker Announcement! 🌟 Introducing our esteemed opening keynote speaker for GenScript's 4th Annual Gene & Cell Engineering Virtual Summit, Dr.…
🌟 Speaker Announcement! 🌟 Introducing our esteemed opening keynote speaker for GenScript's 4th Annual Gene & Cell Engineering Virtual Summit, Dr.…
Liked by Stefano Gullà
-
Oggi si chiude un cerchio grande. Su questo lungomare ci sono cresciuto, ci passavamo le ore di educazione fisica al liceo, organizzavamo i primi…
Oggi si chiude un cerchio grande. Su questo lungomare ci sono cresciuto, ci passavamo le ore di educazione fisica al liceo, organizzavamo i primi…
Liked by Stefano Gullà
-
When sales and marketing align… “The Alignment” is coming 👀 P.S. Joshua Bailey 🦕⛵️ doesn’t know how to use a mic
When sales and marketing align… “The Alignment” is coming 👀 P.S. Joshua Bailey 🦕⛵️ doesn’t know how to use a mic
Liked by Stefano Gullà
-
🚀 Exciting News Alert! 🚀 I'm thrilled to announce the launch of the Malvern Panalytical Zetasizer Advance Pro Sample Assistant! 🎉 This…
🚀 Exciting News Alert! 🚀 I'm thrilled to announce the launch of the Malvern Panalytical Zetasizer Advance Pro Sample Assistant! 🎉 This…
Liked by Stefano Gullà
-
A humanized mouse that mounts mature class-switched, hypermutated and neutralizing antibody responses Many of the more than the 1,600 immune…
A humanized mouse that mounts mature class-switched, hypermutated and neutralizing antibody responses Many of the more than the 1,600 immune…
Liked by Stefano Gullà
-
This recent paper was published in Science Translational Medicine, and entitled “A T cell receptor β chain–directed antibody fusion molecule…
This recent paper was published in Science Translational Medicine, and entitled “A T cell receptor β chain–directed antibody fusion molecule…
Liked by Stefano Gullà
-
Studies on two CD8-targeted IL2 approaches reveal an intriguing mechanism. The transcription factor Tox is involved in T-cell exhaustion, but…
Studies on two CD8-targeted IL2 approaches reveal an intriguing mechanism. The transcription factor Tox is involved in T-cell exhaustion, but…
Liked by Stefano Gullà
-
Prototype #mRNA vaccine for Alzheimer's Disease (AD). Specifically, mRNA encodes 3 copies of the N-terminal region of human Aβ spanning amino acids…
Prototype #mRNA vaccine for Alzheimer's Disease (AD). Specifically, mRNA encodes 3 copies of the N-terminal region of human Aβ spanning amino acids…
Liked by Stefano Gullà
-
A clinical trial began last week for the 5th antibody drug that we discovered in our academic lab at Vanderbilt to get into the clinic. This trial is…
A clinical trial began last week for the 5th antibody drug that we discovered in our academic lab at Vanderbilt to get into the clinic. This trial is…
Liked by Stefano Gullà
-
This most recent open access article was published in Nature Communications, and look interesting to us. Pancreatic ductal adenocarcinoma (PDAC) has…
This most recent open access article was published in Nature Communications, and look interesting to us. Pancreatic ductal adenocarcinoma (PDAC) has…
Liked by Stefano Gullà
-
#Immunotherapy News: Research co-funded by #NIAID found that a type of drug called a JAK inhibitor made a common form of cancer immunotherapy more…
#Immunotherapy News: Research co-funded by #NIAID found that a type of drug called a JAK inhibitor made a common form of cancer immunotherapy more…
Liked by Stefano Gullà
-
AntibodyPlus was introduced by the Chinese Antibody Society (CAS) in 2021 to better reflect the overwhelming growth of antibody-based…
AntibodyPlus was introduced by the Chinese Antibody Society (CAS) in 2021 to better reflect the overwhelming growth of antibody-based…
Liked by Stefano Gullà
Other similar profiles
Explore collaborative articles
We’re unlocking community knowledge in a new way. Experts add insights directly into each article, started with the help of AI.
Explore MoreOthers named Stefano Gullà
2 others named Stefano Gullà are on LinkedIn
See others named Stefano Gullà