Synthesis of ?-aminosuccinimide-containing peptides in Fmoc-based SPPS

Thompson, Phillip E.; Cavallaro, Vittoria; Hearn, Milton Thomas William

doi:10.1007/bf00119766

Cited by 7 publications

(6 citation statements)

References 29 publications

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“…Synthesis of the acetamidomethyl (Acm)‐protected TthL36 polypeptide followed standard solid‐phase peptide synthesis (SPPS) procedures (13,14) with Fmoc‐protected amino acids and the Fmoc‐E‐Rink amide MBHA‐OtBu‐resin (100 µmole eq., Novabiochem, La Jolla, CA). The first amino acid introduced was the Glu residue, which was attached to the resin via its side‐chain.…”

Section: Methodsmentioning

confidence: 99%

The metal binding properties of the CCCH motif of the 50S ribosomal protein L36 from Thermus thermophilus

Boysen

Hearn

2001

The Journal of Peptide Research

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The TthL36 protein of the 50S ribosomal proteins from Thermus thermophilus has been found to contain the rare C(Xaa)2C(Xaa)12C(Xaa)4H (CCCH) sequence motif, a zinc finger binding motif, which for other zinc finger proteins is known to cleave RNA hairpins. In order to investigate the metal-binding properties of this T. thermophilus TthL36 protein, the core 26-mer polypeptide containing this CCCH motif was prepared by solid-phase peptide synthesis methods using Fmoc chemistry, purified by preparative RP-HPLC and characterized by circular dichroism, high-performance capillary zone electrophoresis and electrospray ionization mass spectrometry. Reaction of the acetamidomethyl (Acm)-protected polypeptide with iodine under acidic conditions resulted in the formation of the fully de-protected polypeptide. Of interest, the results demonstrate that the standard Acm-deprotection method with the synthetic TthL36 polypeptide using mercuric acetate in the presence of a large excess of 2-mercaptoethanol resulted in preferential formation of a very stable mercuro-polypeptide complex. The properties of the Acm-deprotected polypeptide in the presence of different metal ions were also investigated by spectroscopic methods. The results confirm that this TthL36 polypeptide containing the CCCH motif binds metal ions with different affinities, namely in the order Co(II)>Hg(II)>Zn(II).

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Section: Methodsmentioning

confidence: 99%

The metal binding properties of the CCCH motif of the 50S ribosomal protein L36 from Thermus thermophilus

Boysen

Hearn

2001

The Journal of Peptide Research

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“…The main side reaction in the synthesis of Aspcontaining peptides is aspartimide formation (Asi: aspartimide; the abbreviation Asu was used in some publications to denote aspartimide [1,2] while the same abbreviation is proposed for α-aminosuberic acid [3]). In order to avoid future confusion we propose the use of Asi as abbreviation, derived from Aspartimide.…”

Section: Introductionmentioning

confidence: 99%

“…Various methods that improve the imide ring formation have been explored, e.g. treatment of the peptide-resin with tertiary amines (10% triethylamine in CH 2 Cl 2 for 15 h) before removal of the solid support with TFMSA, or the Fmoc deprotection with 50% piperidine in DMF (v/v) for 20 min [2].…”

Section: Introductionmentioning

confidence: 99%

α- and β- aspartyl peptide ester formationvia aspartimide ring opening

et al. 2005

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The undesirable reaction of aspartimide formation has been proved to occur under both acid and base conditions in solid-phase peptide synthesis and is dependent on the beta-carboxyl protecting group, the acid or base used during the synthesis, as well as the peptide sequence. The hydrolysis of aspartimide-containing peptides, especially during HPLC purification, yields a mixture of alpha- and beta-aspartyl peptides that can not be purified easily. A previous study demonstrated that treatment of aspartimide-containing peptides with methanol in the presence of 2% diisopropylethylamine in solution leads to alpha- and beta-aspartyl peptide methyl esters. Taking advantage of these results and aiming at elucidating the optimal conditions for aspartimide ring opening, the effect of different types and concentrations of alcohols (primary and secondary) and bases (diisopropylethylamine, collidine, 4-pyrrolidinopyridine, 1-methyl-2-pyrrolidone, piperidine and KCN) was tested at various temperatures and reaction times. The best results were obtained with a combination of a primary alcohol and diisopropylethylamine, while aspartimide ring opening by secondary alcohols occurred only at high temperatures. The optimal conditions were also applied to solid-phase peptide synthesis.

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“…The long treatment of a peptide with strong acids may be detrimental (like in the case of Trpcontaining peptides or' glycopeptides). For the base-catalysed cyclisation of a ]5-benzyl-aspartyl peptide the use of triethylamine (TEA) has been reported [18], but in this case a certain amount of epimerized product may be formed as well, since the aminosuccinimide ring is very prone to basecatalysed racemization [5,19]. To date, the only reliable method that can be used for SPPS as well seems to be that described by Sch6n et al [5], which uses pentafluorophenylester (HOPfp) activation for the aspartyl side chain to achieve the required cyclisation without addition of base.…”

Section: Introductionmentioning

confidence: 99%