Hydrocarbon-Stitched Peptide Agonists of Glucagon-Like Peptide-1 Receptor

Bird, Gregory H.; Fu, Accalia; Escudero, Silvia; Godes, Marina; Opoku-Nsiah, Kwadwo; Wales, Thomas E.; Cameron, Michael D.; Engen, John R.; Danial, Nika N.; Walensky, Loren D.

doi:10.1021/acschembio.0c00308

Cited by 14 publications

(15 citation statements)

References 47 publications

(119 reference statements)

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“…Taken together, HDX provides a high precision read-out of stapling effects on the peptide structure and proteolytic resistance, as recently demonstrated for hydrocarbon-stitched GLP1 peptides. 34 Our SAH-gp41 (638−673) (E) results highlight that when proteolytic resistance is derived more from the sequence-specific staple protection rather than α-helical induction, HDX naturally favors the detection of the conformational rather than the proteolytic changes. Thus, coupling the MS analyses with the study resistance patterns to multiple proteases should prove useful in distinguishing between the effect of hydrocarbon stapling on peptide conformation and the localized protection of vulnerable sequences.…”

Section: ■ Materials and Methodsmentioning

confidence: 87%

“…In vitro proteolysis testing was the most sensitive method for detecting the differential impact of staple variants on the structural stability and, most profoundly, the stability profile was predictive of in vivo activity. 34 However, to our knowledge IM-MS has yet to be leveraged in stapled peptide assessments. Enfuvirtide (Enf) is a 36-residue fusion inhibitor peptide that was approved by the FDA in 2003 for the treatment of HIV-1.…”

Section: ■ Introductionmentioning

confidence: 99%

“…Further, comparative conformational analyses of GLP-1 and variants stabilized by hydrocarbon-stitching revealed that HDX-MS was more effective than circular dichroism at identifying differences in structural stabilization. In vitro proteolysis testing was the most sensitive method for detecting the differential impact of staple variants on the structural stability and, most profoundly, the stability profile was predictive of in vivo activity . However, to our knowledge IM-MS has yet to be leveraged in stapled peptide assessments.…”

Section: Introductionmentioning

confidence: 99%

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Characterizing Native and Hydrocarbon-Stapled Enfuvirtide Conformations with Ion Mobility Mass Spectrometry and Hydrogen–Deuterium Exchange

Stocks

Bird

Walensky

2021

J. Am. Soc. Mass Spectrom.

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The number of approved peptide therapeutics, as well as those in development, has been increasing in recent years. Frequently, the biological activity of such peptides is elicited through the adoption of secondary structural elements upon interaction with their cellular target. However, many therapeutic peptides are unstructured in solution and accordingly exhibit a poor bioavailability due to rapid proteolysis in vivo. To combat this degradation, numerous naturally occurring peptides with therapeutic properties contain stabilizing features, such as N-to-C cyclization or disulfide bonds. Recently, hydrocarbon stapling via non-native amino acid substitution followed by ring-closing metathesis has been shown to induce a dramatic stabilization of α-helical peptides. Identifying the ideal staple location along the peptide backbone is a critical developmental step, and methods to streamline this optimization are needed. Mass spectrometry-based methods such as ion mobility (IM) and hydrogen–deuterium exchange (HDX) can detect multiple discrete peptide conformations, a significant advantage over bulk spectroscopic techniques. In this study we use IM-MS and HDX-MS to demonstrate that the native 36-residue enfuvirtide peptide is highly dynamic in solution and the conformational ensemble populated by stabilized constructs depends heavily on the staple location. Further, our measurements yielded results that correlate well with the average α-helical content measured by circular dichroism. The MS-based approaches described herein represent sensitive and potentially high-throughput methods for characterizing and identifying optimally stapled peptides.

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Section: ■ Materials and Methodsmentioning

confidence: 87%

Section: ■ Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Characterizing Native and Hydrocarbon-Stapled Enfuvirtide Conformations with Ion Mobility Mass Spectrometry and Hydrogen–Deuterium Exchange

Stocks

Bird

Walensky

2021

J. Am. Soc. Mass Spectrom.

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“…Eluant was directed into a Waters Synapt G2si mass spectrometer operated in TOF-only mode for mass analysis. Data were analyzed as described 22 , 40 . All mass spectra were processed manually using MagTran.…”

Section: Methodsmentioning

confidence: 99%

Structural basis for defective membrane targeting of mutant enzyme in human VLCAD deficiency

et al. 2022

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Very long-chain acyl-CoA dehydrogenase (VLCAD) is an inner mitochondrial membrane enzyme that catalyzes the first and rate-limiting step of long-chain fatty acid oxidation. Point mutations in human VLCAD can produce an inborn error of metabolism called VLCAD deficiency that can lead to severe pathophysiologic consequences, including cardiomyopathy, hypoglycemia, and rhabdomyolysis. Discrete mutations in a structurally-uncharacterized C-terminal domain region of VLCAD cause enzymatic deficiency by an incompletely defined mechanism. Here, we conducted a structure-function study, incorporating X-ray crystallography, hydrogen-deuterium exchange mass spectrometry, computational modeling, and biochemical analyses, to characterize a specific membrane interaction defect of full-length, human VLCAD bearing the clinically-observed mutations, A450P or L462P. By disrupting a predicted α-helical hairpin, these mutations either partially or completely impair direct interaction with the membrane itself. Thus, our data support a structural basis for VLCAD deficiency in patients with discrete mutations in an α-helical membrane-binding motif, resulting in pathologic enzyme mislocalization.

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“…It has been widely used for the diagnosis and treatment of T2DM. Recent studies have suggested that GLP-1 can inhibit bone resorption by promoting the secretion of calcitonin[ 3 ]. Matrix Gla protein (MGP) is a circulating protein related to vitamin K that can inhibit calcium and phosphorus deposition and play a role in the regulation of bone metabolism[ 4 ].…”

Section: Introductionmentioning

confidence: 99%

Significance of serum glucagon-like peptide-1 and matrix Gla protein levels in patients with diabetes and osteoporosis

Xie¹,

Zhang²,

Hu³

et al. 2022

WJCC

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BACKGROUND Osteoporosis is a systemic bone disease characterized by decreased bone mass, impaired bone mass, and reduced bone strength that leads to increased bone fragility and fracture. Type 2 diabetes mellitus (T2DM) complicated with osteoporosis is a common systemic metabolic bone disease, and reduced bone mass and bone strength are considered the main clinical features; however, the pathogenesis of this disease has not been fully clarified. Its occurrence is considered related to sex, age, and genetic factors. There are many risk factors for diabetes complicated with osteoporosis. Therefore, exploring these risk factors will help prevent it. AIM To investigate the relationships among serum glucagon-like peptide-1 (GLP-1) levels, matrix Gla protein (MGP) levels, and diabetes with osteoporosis. METHODS Sixty patients with T2DM complicated with osteoporosis confirmed by the endocrinology department of our hospital were selected as the case group. Sixty T2DM patients with bone loss were selected as the control group. Sixty healthy participants were selected as the healthy group. The general data, bone mineral density index, and bone metabolic markers of the three groups were compared. The relationships among GLP-1 levels, MGP levels, and the bone mineral density index of the case group were analyzed using linear correlation analysis and a logistic regression model. RESULTS Differences in sex, smoking, and drinking among the case group, control group, and healthy group were not statistically significant ( P > 0.05). The mean age of the case group was older than those of the control and healthy groups ( P < 0.05). The body mass index, fasting plasma glucose level, HbA1c level, hypertension rate, and coronary heart disease rate of the case and control groups were higher than those of the healthy group ( P < 0.05). The serum GLP-1 and MGP levels of the case group were lower than those of the control and healthy groups; these differences were statistically significant ( P < 0.05). The serum GLP-1 and MGP levels of the control group were lower than those of the healthy group; these differences were statistically significant ( P < 0.05). The serum GLP-1 and MGP levels of the case group were significantly positively correlated with the bone mineral density values of the hip and lumbar spine ( P < 0.05). The results of the logistic regression model showed that age and duration of diabetes were independent risk factors for osteoporosis in diabetic patients ( P < 0.05) and that increased GLP-1 and MGP values were protective factors against osteoporosis in diabetic patients ( P < 0.05). CONCLUSION Serum GLP-1 and MGP levels of diabetic patients with osteoporosis were significantly ...

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Hydrocarbon-Stitched Peptide Agonists of Glucagon-Like Peptide-1 Receptor

Cited by 14 publications

References 47 publications

Characterizing Native and Hydrocarbon-Stapled Enfuvirtide Conformations with Ion Mobility Mass Spectrometry and Hydrogen–Deuterium Exchange

Characterizing Native and Hydrocarbon-Stapled Enfuvirtide Conformations with Ion Mobility Mass Spectrometry and Hydrogen–Deuterium Exchange

Structural basis for defective membrane targeting of mutant enzyme in human VLCAD deficiency

Significance of serum glucagon-like peptide-1 and matrix Gla protein levels in patients with diabetes and osteoporosis

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