Reporter Affinity Labels:  An <i>o</i>-Phthalaldehyde Derivative of β-Naltrexamine as a Fluorogenic Ligand for Opioid Receptors

Bourdonnec, Bertrand Le; Kouhen, Rachid El; Lunzer, Mary M.; Law, Ping Yee; Loh, Horace H.; Portoghese, Philip S.

doi:10.1021/jm000138s

Cited by 22 publications

(21 citation statements)

References 22 publications

(23 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Owing to its rapidity and robustness, the Roth method has gained wide popularity to derivatize AA samples for chromatographic analysis. In addition, it has found usefulness to selectively modify Lys and Cys residues of proteins 42 , 43 . For example, Porttoghese developed an OPA-β-funaltrexamine conjugate as a fluorogenic affinity-labeling ligand of opioid receptors 42 .…”

Section: Resultsmentioning

confidence: 99%

Extendable stapling of unprotected peptides by crosslinking two amines with o-phthalaldehyde

Wang

Chen

et al. 2022

Nat Commun

View full text Add to dashboard Cite

Peptide modification methods that do not rely on the cysteine residue are underdeveloped, and their development could greatly expand the current toolbox for peptide chemistry. During the course of preliminary investigations into the classical ortho-phthalaldehyde (OPA)-amine-thiol condensation reaction, we found that in the absence of thiol, OPA readily condenses with two primary alkyl amines to form a class of underexplored isoindolin-1-imine compounds under mild aqueous conditions. From the intramolecular version of this OPA-2amines reaction, an efficient and selective methodology using mild reaction conditions has been developed for stapling unprotected peptides via crosslinking of two amino groups in both an end-to-side and side-to-side fashion. The stapling method is superfast and broadly applicable for various peptide substrates with the reacting amino groups separated by a wide range of different amino acid units. The macrocyclization reactions of selected substrates are completed within 10 seconds at 5 mM concentration and within 2 minutes at 50 μM concentration. Importantly, the resulting cyclized peptides with an isoindolinimine linkage can be extended in a one-pot sequential addition manner with several different electron-deficient π electrophiles, thereby generating more complex structures.

show abstract

Section: Resultsmentioning

confidence: 99%

Extendable stapling of unprotected peptides by crosslinking two amines with o-phthalaldehyde

Wang

Chen

et al. 2022

Nat Commun

View full text Add to dashboard Cite

show abstract

“…m ‐Phenylsulfonyl fluoride group ( m SF) was expected to display more selective protein labeling than the p ‐isomer ( p SF), which has attenuated reactivity [18] . Bromoacetyl, chloroacetyl and monomethyl fumarate groups have been used as warheads in affinity labeling probes to label the thiol moiety of cysteine residues at enzyme active sites [19–21] . Arylazide group was also included to evaluate the reactivity of multivalent electrophilic groups in comparison [9] .…”

Section: Methodsmentioning

confidence: 99%

Reactivity Analysis of New Multivalent Electrophilic Probes for Affinity Labeling of Carbohydrate Binding Proteins

Kamoshita¹,

Matsui²,

Suto³

et al. 2021

ChemBioChem

View full text Add to dashboard Cite

We have designed and synthesized six different multivalent electrophiles as carbohydrate affinity labeling probes. Evaluation of the reactivity of the electrophiles against peanut agglutinin (PNA) and Ricinus communis agglutinin (RCA) showed that p‐ and m‐aryl sulfonyl fluoride are effective protein reactive groups that label carbohydrate binding lectins in a ligand‐dependent fashion at a nanomolar probe concentration. Analysis of the selectivity of affinity labeling in the presence of excess BSA as a nonspecific protein indicated that m‐arylsulfonyl fluoride is a more selective protein‐reactive group, albeit with attenuated reactivity. Further analysis showed that the labeling efficiency of the multivalent electrophilic probes can be improved by employing reaction conditions involving 25 °C instead of typically employed 4 °C. Both isomers of arylsulfonyl fluoride groups together represent promising affinity labels for target identification studies that could serve as more efficient alternatives to photoreactive groups.

show abstract

“…These ligands include β‐chlornaltrexamine (β‐CNA),3 β‐funaltrexamine (β‐FNA),4 fentanyl isothiocyanate (FIT),5 cis ‐(+)‐3‐methylfentanyl isothiocyanate (SUPERFIT),6 and naltrindole isothiocyanate (NTII) 7. Recently, “reporter” affinity labels, which contain an o ‐phthalaldehyde moiety, have been developed that react with lysine and cysteine residues in the receptor to generate an isoindole fluorophore 8–10…”

Section: Introductionmentioning

confidence: 99%

Synthesis and evaluation of derivatives of leucine enkephalin as potential affinity labels for δ opioid receptors

2003

View full text Add to dashboard Cite

As part of an effort to develop peptide-based affinity labels for opioid receptors, [Leu(5)]enkephalin (LeuEnk) and DTLET (Tyr-D-Thr-Gly-Phe-Leu-Thr), potent agonists for delta receptors, were selected as the parent peptides for further modification. The affinity label derivatives were prepared using standard Fmoc solid-phase peptide synthesis in conjunction with Fmoc-Phe(p-NHAlloc) (Fmoc: 9-flourenylmethoxycarbonyl;) and selective modification of the p-amino group on this residue. The electrophilic isothiocyanate and bromoacetamide groups were introduced into the para position of Phe(4); the corresponding free amine-containing peptides were also prepared for comparison. The pure peptides were evaluated in radioligand binding assays using Chinese hamster ovary (CHO) cells expressing delta and micro opioid receptors. Modification of Phe(4) in LeuEnk and DTLET significantly decreased delta-receptor binding affinity (40 to >2,000-fold). Among the synthesized analogues, [Phe(p-NH(2))(4)]DTLET showed the highest delta-receptor binding affinity (IC(50) = 39 nM) and enhanced selectivity for delta receptors compared to DTLET while other derivatives exhibited much lower delta receptor affinity. The differences in affinities between the two series of analogues and between the derivatives of LeuEnk and N,N-dibenzyl[Leu(5)]Enk reported previously suggest subtle differences in interactions of Phe(4) with delta receptors depending on other modifications in the sequences.

show abstract

Reporter Affinity Labels: An o-Phthalaldehyde Derivative of β-Naltrexamine as a Fluorogenic Ligand for Opioid Receptors

Cited by 22 publications

References 22 publications

Extendable stapling of unprotected peptides by crosslinking two amines with o-phthalaldehyde

Extendable stapling of unprotected peptides by crosslinking two amines with o-phthalaldehyde

Reactivity Analysis of New Multivalent Electrophilic Probes for Affinity Labeling of Carbohydrate Binding Proteins

Synthesis and evaluation of derivatives of leucine enkephalin as potential affinity labels for δ opioid receptors

Contact Info

Product

Resources

About