Overview of Solid Phase Synthesis of “Difficult Peptide” Sequences

Tickler, Anna K.; Wade, John D.

doi:10.1002/0471140864.ps1808s50

Cited by 35 publications

(39 citation statements)

References 26 publications

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“…Successful peptide synthesis is known to technically depend on its length or amino acid sequence (23). Because of the technical difficulty associated with synthesizing the full-length SAA3 peptide without the signal sequence (103 mer), we attempted to shorten the SAA3 peptide sequence while retaining its biological activity.…”

Section: Resultsmentioning

confidence: 99%

Serum Amyloid A3 Binds MD-2 To Activate p38 and NF-κB Pathways in a MyD88-Dependent Manner

Deguchi

Tomita

Omori

et al. 2013

The Journal of Immunology

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Serum amyloid A (SAA) 3 is a major component of the acute phase of inflammation. We previously reported that SAA3 served as an endogenous peptide ligand for TLR4 to facilitate lung metastasis. Because these experiments were performed with SAA3 recombinant proteins purified from Escherichia coli or mammalian cells, we could not rule out the possibility of LPS contamination. In this study, we used SAA3 synthetic peptides to eliminate the presence of LPS in SAA3. We found that the SAA3 synthetic peptide (aa 20–86) (20–86) stimulated cell migration and activated p38 in a manner dependent on TLR4, MD-2, and MyD88. SAA3 (20–86) also activated NF-κB and Rho small GTPase. Using surface plasmon resonance analysis, the binding constant KD values between SAA3 (20–86) or SAA3 (43–57) and TLR4/MD-2 protein highly purified by the baculovirus system were 2.2 and 30 μM, respectively. FLAG-tagged SAA3 tightly bound to protein A–tagged MD-2, but not to TLR4 in baculovirus coinfection experiments. Although SAA3 (20–86) caused a low, but appreciable level of endocytosis in TLR4, it induced the upregulation of both IL-6 and TNF-α, but not IFN-β1. An i.v. injection of SAA3 (43–57) induced the lung recruitment of CD11b+Gr-1+ cells at an estimated serum concentration around its KD value toward TLR4/MD-2. Taken together, these results suggest that SAA3 directly binds MD-2 and activates the MyD88-dependent TLR4/MD-2 pathway.

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Section: Resultsmentioning

confidence: 99%

Serum Amyloid A3 Binds MD-2 To Activate p38 and NF-κB Pathways in a MyD88-Dependent Manner

Deguchi

Tomita

Omori

et al. 2013

The Journal of Immunology

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“…Since Merrifield invented the technology in 1963, SPPS has become the primary approach for the routine preparation of peptides [28][29][30][31][32]. The general scheme of solid phase peptide synthesis is illustrated in Fig.…”

Section: Synthesis Of Peptide Fragmentsmentioning

confidence: 99%

New Methods for Chemical Protein Synthesis

Guan

Chaffey

Zeng

2014

Topics in Current Chemistry

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Chemical protein synthesis is a useful tool to generate pure proteins which are otherwise difficult to obtain in sufficient amounts for structure and property analysis. Additionally, because of the precise and flexible nature of chemical synthesis, it allows for controllable variation of protein sequences, which is valuable for understanding the relationships between protein structure and function. Despite the usefulness of chemical protein synthesis, it has not been widely adopted as a tool for protein characterization, mainly because of the lack of general and efficient methods for the preparation and coupling of peptide fragments and for the folding of polypeptide chains. To address these issues, many new methods have recently been developed in the areas of solid-phase peptide synthesis, peptide fragment assembly, and protein folding. Here we review these recent technological advances and highlight the gaps needing to be addressed in future research.

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“…[86] Recombinant expression of this protein is limited due to the high number of hydroxyproline residues.…”

Section: Native Chemical Ligationmentioning

confidence: 99%