PURE™ Peptides From CRB

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Case Study

Post-translational modification study using acetylated and methylated peptides

PURE™ Peptides

CRB offers a range of peptide service categories providing every type of peptide request possible

Histone H3 Peptide

Ease of synthesis of histone peptides

Using acetylated and methylated peptides synthesised at Cambridge Research Biochemicals it has been demonstrated that there is an interplay between NF-κB subunit Re1A acetylation and methylation.

Post-translational modifications of the Re1A subunit of NF-κB play a key role in controlling its nuclear activity. These modifications include acetylation and methylation. In this work chemically synthesised peptides with and without acetylation and methylation modifications have been used to establish that acetylation of lysine 310 of Re1A impairs the Set9-mediated methylation of lysines 314 and 315, which is important for the ubiquitination and degradation of chromatin–associated Re1A. NF-κB plays an important role in regulating inflammatory responses, apoptosis, cell proliferation and differentiation and tumorigenesis.

Standard peptides are made up of naturally occurring L-amino acids (of which there are twenty) and have a length of 6-25 residues. We offer two purity standards: >80% and >95% by HPLC and can synthesise laboratory-scale amounts from 1mg – gram scale quantities. Turnaround is 2-3 weeks.

Modified peptides include cyclisation, phosphorylation, sulphation, methylation, glycosylation and N/C termini modification.

Extensive Capability

	Routine peptides - 1 mg-10 g
	Single peptides to large arrays
	Cyclic peptides - disulphide and head-to-tail, side-chain to side-chain or backbone lactam bridged
	Post-translational modifications - phosphorylated, sulphated, methylated & glycosylated peptides
	C-terminal modifications - secondary amides, alcohols, aldehydes, esters, AMCs, pNAs

Peptide Applications

Within the field of biology chemically synthesised peptides are a very powerful research tool. Peptides can be used in wide-ranging applications both in-vitro and in-vivo to investigate biological pathways and hence are an integral part of the drug discovery process. Some typical applications our peptides are used for are:

	Production of anti-protein antibodies where a peptide is used as a surrogate for the protein
	Enzyme activity studies in which a peptide substrate is used to investigate actions of enzymes such as kinases which are critically important in cancer research
	Binding studies in which a fluorescently labelled or radiolabelled peptide is used to study binding of the peptide ligand to proteins
	Study of post-translational modifications such as phosphorylation and methylation where these modifications can be chemically included within a peptide

Quality

	Purity: >95% by HPLC
	Peptide identity confirmed by mass spectrometry (ESI or MALDI-TOF)
	In-process control of the synthesis of complex peptides by mass spectrometry and HPLC monitoring

Customer Support

At CRB we pride ourselves on the quality of our peptides and our customer focussed service:

	Detailed consultation on peptide design
	Superior building block and synthesis strategies
	Custom chemical synthesis of unusual reagents
	Regular progress updates
	Custom aliquotting upon request
	Excess peptide retained in stock
	Peptide Inventory dating back to 1980

Acetyl-Leu-Ala-Thr-Lys-Ala-Ala-(ADMA)-Lys(Me)₃
-Ser-Ala-Pro-Ala-Thr-Gly-Gly-Val-Lys-Lys-Pro
-His-Arg-Ahx-Lys(Biotin)-NH₂

Post-translational modifications of Histones (Acetylation, methylation, phosphorylation…) alter their interaction with DNA and act in diverse biological processes such as gene regulation, DNA repair etc. With the commercial introduction of methylated arginine (mono-, assymmetric di- and symmetric di-) and lysine (mono-, di- and tri-), synthesis of histone peptides has become more straightforward than ever. C-terminal biotinylated H3 (20-40) was synthesised using minimal amounts of assymmetric dimethylarginine and trimethyl lysine.

Unmodified, K-acetylated and K-monomethylated versions of TYETFKSIMKKSPFSGPTDP-acid
Functional Interplay between Acetylation and Methylation of the Re1A Subunit of NF-κB
X-D. Yang, E. Tajkhorshid & L-F. Chen. University of Illinois
Molecular & Cellular Biology 2010, 30, 9, 2170-2180

MEVGWYRSPFSRVVHLYRNGK-acid Myelin Oligodendrocyte Peptide (MOG 35-55)
Modulation of experimental autoimmune encephalomyletis by endogenous Annexin A1
N. Paschalidis, A. J. Iqbal, F. Maione, E. G. Wood, M. Perreti, R. J. Flower & F. D’Acquisto. Queen Mary University of London
Journal of Neuroinflammation 2009, 6:33

Multiple peptides with N-terminal biotin up to 35 residues in length
Two modules in the BRC repeats of BRCA2 mediate structural and functional interactions with the RAD51 recombinase
E. Rajendra & A. R. Venkitaraman. Medical Research Council Cancer Cell Unit
Nucleic Acids Research, 2009, 1-15

Should you require a quotation from any of our custom services, we have a clear cut and easy system that enables you to do so. Follow these three simple steps:

Go to: www.crbdiscovery.com/quotations

Simply click on the relevant tab and register your details.

Once issued with a password you can then log on to our quotation system using your customer account log-in and a quotation will be issued within 24 hours.

At Cambridge Research Biochemicals we have a fast-track quotation system that tailors your requirements, gives you all the information you need and a specific price within 24 hours. You are also given a personal contact so that if you have any further questions you can directly contact one of our approachable representatives who has the technical knowledge to assist you.