DABCYL succinimidyl ester [4-((4-(Dimethylamino)phenyl)azo)benzoic acid, succinimidyl ester] CAS 146998-31-4

Methionine aminopeptidase (MetAP) catalyzes the hydrolytic cleavage of the N-terminal methionine from newly synthesized polypeptides. The extent of removal of methionyl from a protein is dictated by its N-terminal peptide sequence. Earlier studies revealed that MetAPs require amino acids containing small side chains (e.g., Gly, Ala, Ser, Cys, Pro, Thr, and Val) as the P1′ residue, but their specificity at positions P2′ and beyond remains incompletely defined. In this work, the substrate specificities of <em>Escherichia coli</em> MetAP1, human MetAP1, and human MetAP2 were systematically profiled by screening against a combinatorial peptide library and kinetic analysis of individually synthesized peptide substrates. Our results show that although all three enzymes require small residues at the P1′ position, they have differential tolerance for Val and Thr at this position. The catalytic activity of human MetAP2 toward Met-Val peptides is consistently 2 orders of magnitude higher than that of MetAP1, suggesting that MetAP2 is responsible for processing proteins containing N-terminal Met-Val and Met-Thr sequences <em>in vivo</em>. At positions P2′−P5′, all three MetAPs have broad specificity but are poorly active toward peptides containing a proline at the P2′ position. In addition, the human MetAPs disfavor acidic residues at the P2′−P5′ positions. The specificity data have allowed us to formulate a simple set of rules that can reliably predict the N-terminal processing of <em>E. coli</em> and human proteins. Source: <strong>Protein N-Terminal Processing: Substrate Specificity of Escherichia coli and Human Methionine Aminopeptidases</strong> by Xiao et al., <em>ACS Publications</em>, June 2010.

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Physical properties

Molecular weight	366.37
Solvent	DMSO

Spectral properties

Absorbance (nm)	454
Correction Factor (280 nm)	0.516

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12352200

Overview SDS Protocol

CAS

146998-31-4

Molecular weight

366.37

Absorbance (nm)

454

Correction Factor (280 nm)

0.516

DABCYL, SE is the amino-reactive form of DABCYL, and widely used to prepare a variety of FRET-based probes that contain DABCYL. DABCYL is one of the most popular acceptors for developing FRET-based nucleic acid probes and protease substrates.

Calculators

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of DABCYL succinimidyl ester [4-((4-(Dimethylamino)phenyl)azo)benzoic acid, succinimidyl ester] *CAS 146998-31-4* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

	0.1 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	272.948 µL	1.365 mL	2.729 mL	13.647 mL	27.295 mL
5 mM	54.59 µL	272.948 µL	545.896 µL	2.729 mL	5.459 mL
10 mM	27.295 µL	136.474 µL	272.948 µL	1.365 mL	2.729 mL

Molarity calculator

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Spectrum

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Spectral properties

Absorbance (nm)	454
Correction Factor (280 nm)	0.516

Images

Figure 1. Methionine aminopeptidase (MetAP) catalyzes the hydrolytic cleavage of the N-terminal methionine from newly synthesized polypeptides. The extent of removal of methionyl from a protein is dictated by its N-terminal peptide sequence. Earlier studies revealed that MetAPs require amino acids containing small side chains (e.g., Gly, Ala, Ser, Cys, Pro, Thr, and Val) as the P1′ residue, but their specificity at positions P2′ and beyond remains incompletely defined. In this work, the substrate specificities of Escherichia coli MetAP1, human MetAP1, and human MetAP2 were systematically profiled by screening against a combinatorial peptide library and kinetic analysis of individually synthesized peptide substrates. Our results show that although all three enzymes require small residues at the P1′ position, they have differential tolerance for Val and Thr at this position. The catalytic activity of human MetAP2 toward Met-Val peptides is consistently 2 orders of magnitude higher than that of MetAP1, suggesting that MetAP2 is responsible for processing proteins containing N-terminal Met-Val and Met-Thr sequences in vivo. At positions P2′−P5′, all three MetAPs have broad specificity but are poorly active toward peptides containing a proline at the P2′ position. In addition, the human MetAPs disfavor acidic residues at the P2′−P5′ positions. The specificity data have allowed us to formulate a simple set of rules that can reliably predict the N-terminal processing of E. coli and human proteins. Source: Protein N-Terminal Processing: Substrate Specificity of Escherichia coli and Human Methionine Aminopeptidases by Xiao et al., ACS Publications, June 2010.

Figure 2. Chemical structure for DABCYL succinimidyl ester [4-((4-(Dimethylamino)phenyl)azo)benzoic acid, succinimidyl ester] *CAS 146998-31-4*

Citations

View all 1 citations: Citation Explorer

Protein N-terminal processing: substrate specificity of Escherichia coli and human methionine aminopeptidases
Authors: Xiao, Qing and Zhang, Feiran and Nacev, Benjamin A and Liu, Jun O and Pei, Dehua
Journal: Biochemistry (2010): 5588--5599

References

View all 34 references: Citation Explorer

Recombinant expression and partial characterization of an active soluble histo-aspartic protease from Plasmodium falciparum
Authors: Xiao H, Sinkovits AF, Bryksa BC, Ogawa M, Yada RY.
Journal: Protein Expr Purif (2006): 88

Internally quenched peptides for the study of lysostaphin: An antimicrobial protease that kills Staphylococcus aureus
Authors: Warfield R, Bardelang P, Saunders H, Chan WC, Penfold C, James R, Thomas NR.
Journal: Org Biomol Chem (2006): 3626

A quenched fluorescent dipeptide for assaying dispase- and thermolysin-like proteases
Authors: Weimer S, Oertel K, Fuchsbauer HL.
Journal: Anal Biochem (2006): 110

Characterization and inhibition of SARS-coronavirus main protease
Authors: Liang PH., undefined
Journal: Curr Top Med Chem (2006): 361

Fluorogenic peptide substrates containing benzoxazol-5-yl-alanine derivatives for kinetic assay of cysteine proteases
Authors: Szabelski M, Rogiewicz M, Wiczk W.
Journal: Anal Biochem (2005): 20

ADAM33 enzyme properties and substrate specificity
Authors: Zou J, Zhang R, Zhu F, Liu J, Madison V, Uml and SP., undefined
Journal: Biochemistry (2005): 4247

Enzymatic activity characterization of SARS coronavirus 3C-like protease by fluorescence resonance energy transfer technique
Authors: Chen S, Chen LL, Luo HB, Sun T, Chen J, Ye F, Cai JH, Shen JK, Shen X, Jiang HL.
Journal: Acta Pharmacol Sin (2005): 99

Fluorogenic peptide substrates for carboxydipeptidase activity of cathepsin B
Authors: Stachowiak K, Tokmina M, Karpinska A, Sosnowska R, Wiczk W.
Journal: Acta Biochim Pol (2004): 81

Synthesis of poly(ethylene glycol)-based saquinavir prodrug conjugates and assessment of release and anti-HIV-1 bioactivity using a novel protease inhibition assay
Authors: Gunaseelan S, Debrah O, Wan L, Leibowitz MJ, Rabson AB, Stein S, Sinko PJ.
Journal: Bioconjug Chem (2004): 1322

Highly sensitive intramolecularly quenched fluorogenic substrates for renin based on the combination of L-2-amino-3-(7-methoxy-4-coumaryl)propionic acid with 2,4-dinitrophenyl groups at various positions
Authors: Paschalidou K, Neumann U, Gerhartz B, Tzougraki C.
Journal: Biochem J (2004): 1031