Metal‐Chelating Amino Acids As Building Blocks For Synthetic Receptors Sensing Metal Ions And Histidine‐Tagged Proteins

Hutschenreiter, Silke; Neumann, Lars; Rädler, Ulf; Schmitt, Lutz; Tampé, Robert

doi:10.1002/cbic.200200455

Cited by 34 publications

(25 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Other authors subsequently published the procedure for the synthesis of Ada n -containing peptides using Fmoc-Ada n -OH protected synthons. [18,19] However, the Fmoc group was introduced at the end of the synthetic sequence by using a temporary protection of the amine group. As a result, lengthy procedures involving multiple protection/deprotection steps were reported.…”

Section: Resultsmentioning

confidence: 99%

“…[16] Among metal-chelating amino acid side-chains, aminodiacetate groups have attracted our attention. [17][18][19] The corresponding residues are referred to as Ada n (n being the length of the alkyl chain separating the peptide backbone from the aminodiacetate nitrogen). They have been shown to stabilize helical structures in synthetic peptides through divalent-metal-ion coordination.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Lanthanide(III) Complexes with Two Hexapeptides Incorporating Unnatural Chelating Amino Acids: Secondary Structure and Stability

Cisnetti

Gateau

Lebrun

et al. 2009

Chemistry A European J

View full text Add to dashboard Cite

Unnatural metal-chelating amino acids bearing aminodiacetate side-chains have been introduced into two hexapeptides to obtain efficient lanthanide-binding peptides. The synthesis of the enantiopure Fmoc-Ada(n)(tBu)2-OH synthons is described with overall yields of 32 and 50% for n=2 and n=3 side-chain carbon atoms, respectively. The two peptides AcWAda(n)PGAda(n)GNH2 (Pn) were synthesized from the protected synthons by standard solid-phase peptide synthesis. Studies of the lanthanide complexes of the two peptides Pn by luminescence titrations, mass spectrometry, circular dichroism, and solution NMR spectroscopy demonstrate that the Ada(n) chain length has a dramatic effect on the complexation properties. Indeed, the flexible compound P3 forms a mononuclear complex of moderate stability (beta11=10(9.9)), which tends to transform into a binuclear species in the presence of excess of the metal ion. Interestingly, the more compact peptide P2 provides stable Ln3+ complexes with the exclusive formation of the mononuclear LnP2 adduct. The stability constant of TbP2 is two orders of magnitude higher (beta11=10(12.1)) than that measured for P3. The 800 MHz NMR spectrum of the La3+ complex of P2 evidences a well-defined type II beta-turn as well as a hydrophobic Trp(indole)-Pro interaction. These interactions exemplify the non-innocent character of the peptide spacer in the complex LaP2 as well as the role of a peptide secondary structure in the stabilization of metal complexes.

show abstract

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Lanthanide(III) Complexes with Two Hexapeptides Incorporating Unnatural Chelating Amino Acids: Secondary Structure and Stability

Cisnetti

Gateau

Lebrun

et al. 2009

Chemistry A European J

View full text Add to dashboard Cite

show abstract

“…The synthesis of Fmoc-Lys(IDA)(OtBu)-OH has been previously reported [19]. The synthesis of peptide 1 is shown as a representative example in Scheme 1 using Fmoc-Lys(IDA(OtBu))-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Ile-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Tyr(H 2 PO 3 )-OH, and Fmoc-Lys(IDA(OtBu))-OH, respectively, in coupling reactions with the resin.…”

Section: 0 Results and Disucssionmentioning

confidence: 99%

“…The synthesis of Fmoc-Lys(IDA)-OH has been previously described [19]. The preparation of peptides containing the IDA moiety was carried out according to the previously described procedure [15] using Fmoc-Lys(IDA(OtBu))-OH as the building block.…”

Section: 0 Materials and Methodsmentioning

confidence: 99%

Synthesis and evaluation of phosphopeptides containing iminodiacetate groups as binding ligands of the Src SH2 domain

Schuler

Ahmadibeni

et al. 2009

Bioorganic Chemistry

View full text Add to dashboard Cite

Phosphopeptide pTyr-Glu-Glu-Ile (pYEEI) has been introduced as an optimal Src SH2 domain ligand. Peptides, Ac-K(IDA)pYEEIEK(IDA) (1), Ac-KpYEEIEK (2), Ac-K(IDA)pYEEIEK (3), and Ac-KpYEEIEK(IDA) (4), containing 0-2 iminodiacetate (IDA) groups at the N-and C-terminal lysine residues were synthesized and evaluated as the Src SH2 domain binding ligands. Fluorescence polarization assays showed that peptide 1 had a higher binding affinity (K d = 0.6 µM) to the Src SH2 domain when compared with Ac-pYEEI (K d = 1.7 µM), an optimal Src SH2 domain ligand, and peptides 2−4 (K d = 2.9-52.7 µM). The binding affinity of peptide 1 to the SH2 domain was reduced by more than two fold (K d = 1.6 µM) upon addition of Ni 2+ (300 µM), possibly due to modest structural effect of Ni 2+ on the protein as shown by circular dichroism experimental results. The binding affinity of 1 was restored in the presence of EDTA (300 µM) (K d = 0.79 µM). These studies suggest that peptides containing IDA groups may be used for designing novel SH2 domain binding ligands. Keywordsphosphopeptides; metal chelation; iminodiacetate group; SH2 domain; UV titration; fluorescence polarization; circular dichroism IntroductionSrc, one of the first studied non-receptor tyrosine kinases, has been implicated in the genesis and progression of multiple types of human diseases including colon, breast, lung cancers, osteoporosis, and inflammation-mediated bone loss [1,2]. Src tyrosine kinase contains three major domains, in the order from N-to C-terminal, two regulatory Src homology (SH) domains (SH3 and SH2 domains), and a kinase domain. The Src SH2 domain is a relatively small protein module of approximately 100 amino acids and has extraordinary ability to recognize specifically sequences containing phosphotyrosine residue (pTyr), thereby facilitating phosphorylation-dependent protein-protein interactions that result in signal transduction process [3]. Short peptide inhibitors capable of disrupting these interactions have been designed as useful tools in the mechanistic studies of the Src SH2 domain interactions and as potential NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript inhibitors for further pharmaceutical development [4−8]. The Src SH2 domain preferentially binds peptides with the sequence pTyr-Glu-Glu-Ile (pYEEI) with high affinity [9].The use of conformationally constrained peptides has been shown to be an effective strategy in developing therapeutic peptidic and peptidomimetic agents [10,11]. Constrained peptides are usually more stable towards enzymatic degradations. We previously reported the synthesis and evaluation of a series of covalently bonded conformationally constrained peptide analogues based on the optimal sequence pYEEI as inhibitors of the Src SH2 domain [8]. A number of conformationally constrained peptides showed at least 100-fold fold increase in the binding affinities to the Src SH2 domain relative to pYEEI and the corresponding linear peptides, respectively. The enhancement in binding affinities was a...

show abstract

“…10 À6 m). [19][20][21] This dramatic decrease of the affinity constant has been ascribed to the multivalent interactions offered to multiple mono-NTA binders with polyhistidine tags. [22] Therefore, the design and synthesis of supramolecules bearing several NTA moieties appeared to be a rational strategy for the development of probes displaying a strong affinity for histidinetagged proteins.…”

Section: Introductionmentioning

confidence: 99%

Insight into the Complexation Mode of Bis(nitrilotriacetic acid) (NTA) Ligands with Ni²⁺ Involved in the Labeling of Histidine‐Tagged Proteins

Brellier

Barlaam

Mioskowski

et al. 2009

Chemistry A European J

View full text Add to dashboard Cite

According to literature reports and our own findings, the binding of new Ni(2+)-preloaded bis(nitrilotriacetic acid) (NTA) ligands with polyhistidine-tagged proteins has been found to be accompanied by a one- to two-order-of-magnitude increase in affinity, compared to the binding of a single Ni(2+)-preloaded NTA moiety. In spite of the introduction of a second NTA chelating group, a cooperative effect that is less than the theoretical maximum has been observed. Herein, we present a rational explanation for the observed stability of the ternary complex involving the postulated bis-NTA-(Ni(2+))(2) species and multivalent polyhistidine tags. We have found that prior to the formation of the ternary complex, the Ni(2+)-preloading step of bis-NTA ligands does not form the expected bis-NTA-(Ni(2+))(2) exclusively. Instead of the major formation of bis-NTA-(Ni(2+))(2) species, it appears that cyclic discrete 1:1 and 2:2 entities are predominantly formed. It is proposed that these species interact upon ring-opening with multivalent histidine tags. The occurrence of this phenomena accounts for the overall one- to two-order-of-magnitude increase in affinity of ternary complexes involving bis-NTA ligands.

show abstract

Metal‐Chelating Amino Acids As Building Blocks For Synthetic Receptors Sensing Metal Ions And Histidine‐Tagged Proteins

Cited by 34 publications

References 35 publications

Lanthanide(III) Complexes with Two Hexapeptides Incorporating Unnatural Chelating Amino Acids: Secondary Structure and Stability

Lanthanide(III) Complexes with Two Hexapeptides Incorporating Unnatural Chelating Amino Acids: Secondary Structure and Stability

Synthesis and evaluation of phosphopeptides containing iminodiacetate groups as binding ligands of the Src SH2 domain

Insight into the Complexation Mode of Bis(nitrilotriacetic acid) (NTA) Ligands with Ni²⁺ Involved in the Labeling of Histidine‐Tagged Proteins

Contact Info

Product

Resources

About

Metal‐Chelating Amino Acids As Building Blocks For Synthetic Receptors Sensing Metal Ions And Histidine‐Tagged Proteins

Cited by 34 publications

References 35 publications

Lanthanide(III) Complexes with Two Hexapeptides Incorporating Unnatural Chelating Amino Acids: Secondary Structure and Stability

Lanthanide(III) Complexes with Two Hexapeptides Incorporating Unnatural Chelating Amino Acids: Secondary Structure and Stability

Synthesis and evaluation of phosphopeptides containing iminodiacetate groups as binding ligands of the Src SH2 domain

Insight into the Complexation Mode of Bis(nitrilotriacetic acid) (NTA) Ligands with Ni2+ Involved in the Labeling of Histidine‐Tagged Proteins

Contact Info

Product

Resources

About

Insight into the Complexation Mode of Bis(nitrilotriacetic acid) (NTA) Ligands with Ni²⁺ Involved in the Labeling of Histidine‐Tagged Proteins