Immobilized Adenosine Nucleotides

Ligand:  ATP  |  dATP  |  ADP  |  AMP  |  cAMP

 
Adenosine triphosphate (ATP)-affinity chromatography has been widely used to purify various ATP-binding proteins such as kinases [1-4], β-and γ-glutamate decarboxylase [5] and chaperones [6-9].

Efficient protein binding strongly depends on and the type of ATP-matrix attachment [4,10-11].

 

Jena Bioscience provides a large selection of ready-to-use ATP affinity chromatography material

ATP Positions

  • ATP immobilized at different positions:
    Adenine base moiety: C-8, N-6
    Ribose moiety: 2'/3'-OH
    Phosphate moiety: γ-Phosphate
  • several types and lengths of linkers
  • different types of chromatography material ranging from bulk material to pre-made columns
 
Try our ATP Affinity Test Kit to find the most suitable material for the purification of your protein

 
 
 
 
 


Linked via the γ-Phosphate

Ligand
Cat.-No.

Attachment Position
Linker Type

Structure

γ-Aminophenyl-ATP
AC-101

γ-Phosphate
10-atom carbon spacer

Immobilized gamma-Aminophenyl-ATP (C10-spacer)

γ-Aminophenyl-ATP
AC-102

γ-Phosphate
without spacer

Immobilized gamma-Aminophenyl-ATP (no spacer)

γ-Amino-octyl-ATP
AC-105

γ-Phosphate
8-atom carbon spacer

Immobilized gamma-Aminooctyl-ATP

γ-Amino-hexyl-ATP
AC-116

γ-Phosphate
6-atom carbon spacer

Immobilized gamma-Aminohexyl-ATP

 
Linked via the Nucleobase

Ligand
Cat.-No.

Attachment Position
Linker Type

Structure

8-Amino-hexyl-ATP*
AC-127

C-8
6-atom carbon spacer

Immobilized 8-Aminooctyl-ATP

8-Amino-butyl-ATP*
AC-128

C-8
4-atom carbon spacer

Immobilized 8-Aminobutyl-ATP

N6-(6-Amino)hexyl-ATP*
AC-129

N-6
6-atom carbon spacer

Immobilized N6-Aminohexyl-ATP

N6-(4-Amino)butyl-ATP*
AC-130

N-6
4-atom carbon spacer

Immobilized N6-Aminobutyl-ATP

 
Linked via the Ribose

Ligand
Cat.-No.

Attachment Position
Linker Type

Structure

2'/3'-EDA-ATP*
AC-131

2'/3'-OH
EDA

Immobilized EDA-ATP

 
* Phosphorous chain may be degraded by NTP/dNTP hydrolyzing enzymes.
 
Selected references:

[1] Jeansonne et al. (2006) A rapid ATP affinity-based purification for the human non-receptor tyrosine kinase c-Src. Protein Expression and Purification 46:240.

[2] Dhillon et al. (2009) The C-terminus of Raf-1 acts as a 14-3-3-dependent activation switch. Cellular Signalling 21 (11):1645.

[3] Mlakar et al. (2006) Citrate Inhibition-Resistant Form of 6-Phosphofructo-1-Kinase from Aspergillus niger. Applied and Environmental Microbiology 72 (7):4515.

[3] Ramadoss et al. (1976) Affinity chromatography of phosphofructokinase. Arch. Biochem. Biophys. 175 (2):487.

[4] Haystead et al. (1983) Gamma-phosphate-linked ATP-sepharose for the affinity purification of protein kinases- rapid purification to homogeneity of skeletal muscle mitogen-activated protein kinase kinase. Eur. J. Biochem. 214 (2):459.

[5] Wu et al. (1984) Binding of ATP to brain glutamate decarboxylase as studied by affinity chromatography. J. Neurochem. 42 (6):1607.

[6] Bendz et al. (2007) Human heat shock protein 70 enhances tumor antigen presentation through complex formation and intracellular antigen delivery without innate immune signaling. J. Biol. Chem. 282 (43):31688.

[7] Place et al. (2001) Temperature interactions of the molecular chaperone Hsc70 from the eurythermal marine goby Gillichthys Mirabilis. Journal of Experimental Biology 204:2675.

[8] Kumaraguru et al. (2000) Involvement of an ATP-Dependent Peptide Chaperone in Cross-Presentation after DNA Immunization. Journal of Immunology 165:750.

[9] Welch et al. (1985) Rapid Purification of Mammalian 70,000-Dalton Stress Proteins: Affinity of the Proteins for Nucleotides. Molecular and Cellular Biology 5 (6):1229.

[10] Jeno et al. (1989) Purification and Charaterization of a 40 S Ribosomal Protein S6 Kinase from Vanadate-stimulated Swiss 3T3 Cells. J. Biol. Chem. 264:1293.

[11] Trayer et al. (1974) Affinity Chromatography of Nicotinamide Nucleotide-Dependent Dehydrogenases on Immobilized Nucleotide Derivates. Biochem. J. 139:609.