Pf1 phage for NMR analysis

Pf1 Nuclear Magnetic Resonance Cosolvent

Filamentous bacteriophage for alignment RNA, DNA, and proteins for measurement of nuclear magnetic resonance dipolar coupling interactions

A B C

Fig.1: 

A. Pf1 phage strain LP11-92 is isolated from wild type Pseudomonas aeruginosa and propagated in the phage free strain LA23-99. 

B. KBr gradient of Pf1 preparation. Phage band is marked by the yellow arrow, membrane fraction is shown by red one. 

C.The electron microscopy picture of the pf1 preparation is kindly provided by Dr. Velta Ose, Biomedicine Research and Study Center, Riga, Latvia.

 

The phage is purified on KBr gradient and purchased in a suspension of 10mM K-phosphate buffer pH7.6, 2mM MgCl2 and 0.05% NaN3 (buffer content may be adjusted as to customer needs).

References on Pf1 mediated tunable alignment of proteins and nucleic acids

  1. Hansen MR, Mueller L, Pardi A. (1998) Tunable alignment of macromolecules by filamentous phage yields dipolar coupling interactions. Nat Struct Biol Dec;5(12):1065-74
  2. Clore, G.M., Starich, M.R., Gronenborn, A.M. (1998) Measurement  of residual dipolar couplings of macromolecules aligned in the  nematic phase of a colloidal suspension of 
    rod-shaped viruses.   J. Am. Chem. Soc. 120 , 10571-10572.
  3. Markus MA, Gerstner RB, Draper DE, Torchia DA.(1999) Refining the overall structure and subdomain orientation of ribosomal protein S4 delta41 with dipolar couplings measured by NMR in uniaxial liquid crystalline phases.J Mol Biol Sep 17;292(2):375-87
  4.  Hansen MR, Hanson P, Pardi A. (2000) Filamentous bacteriophage for aligning RNA, DNA, and proteins for measurement of nuclear magnetic resonance dipolar coupling interactions.Methods Enzymol; 317:220-40
  5.  Ramirez BE, Voloshin ON, Camerini-Otero RD, Bax A.(2000) Solution structure of DinI provides insight into its mode of RecA inactivation. Protein Sci. Nov; 9 (11):2161-9
  6. Zweckstetter M, Bax A.(2001) Characterization of molecular alignment in aqueous suspensions of Pf1 bacteriophage.J Biomol NMR Aug;20(4):365-77
  7. Markus Zweckstetter. Residual dipolar couplings and orientational effects Max-Planck-Institute for Biophysical Chemistry, Göttingen mzwecks@gwdg

 

Tunable alignments

Quadrupolar splittings of the 2H NMR signal of D2O in the presence of different concentrations of Pf1 phage LP11-92, recorded at 25oC. The measurements are done in Karolinska Institute, Department of Medical Biochemistry and Biophysics in the group of Professor Gottfried Otting
Fig3. The relationship between LP11-92 phage concentration and delta/Hz
 
 
 
 
 
 
Fig.2 Raw data of quadrupolar splitting  

 


Technical data

BUFFER CONDITONS USED FOR ALIGNMENT OF MACROMOLECULES

WITH PF1 FILAMENTOUS PHAGE

(compiled from Methods in Enzymology, 2000, Vol. 317, 220-240).

BUFFER CONDITIONS Normalized 2H residual coupling (Hz)
10 mM NaH2PO4, 1.5 mM EDTA, pH 8, ~17 mg/ml Pf1, 0.3 mM IRE RNA 8.2
40 mM NaH2PO4, 0.5 mM EDTA, pH 8, ~13 mg/ml Pf1, 1.8 mM DNA 16-mer 6.0
10 mM NaH2PO4, 100 mM NaCI, pH 6.8, ~6 mg/ml Pf1, 1 mM DNA 10-mer 7.7
10 mM Tris(d11), 0.5 mM EDTA, pH 8, ~22 mg/ml Pf1, 0.9 mM IRE RNA 6.1
10 mM imidazole(d4), 5 mM KCl, 0.5 mM EDTA, pH 6.5, ~22 mg/ml Pf1,1.9 mM apocalmodulin, 7.2
10 mM succinate, 0.1 mM EDTA, pH 5.5, ~19 mg/ml Pf1 8.2 (7.3)
10 mM succinate, 100 mM NaCl, 0.1 mM EDTA, pH 5.5, ~19 mg/ml Pf1 7.8 (5.0)
10 mM succinate, 100 mM NaCl, 2.0 mM MgCl2, 0.1 mM EDTA, pH 5.5, ~20 mg/ml Pf1 6.9 (4.6)
10 mM Tris(d11), 0.1 mM EDTA, pH 8, ~15 mg/ml Pf1 8.8
10 mM Tris(d11), 0.1 mMEDTA, pH 7, ~15 mg/ml Pf1 5.5
10 mM Tris(d11), 0.1 mM EDTA, pH 6, ~15 mg/ml Pf1 4.4

The residual quadrupole coupling constants for the D2O signal were measured by 1-D2H spectra. Because all buffer conditions did not have the same Pf1 concentration, the values given are normalized to Pf1 phage concentration of 10 mg/ml. Values in parentheses indicate that the sample was prepared by dialysis, whereas the other samples were prepared by pelleting the phage in the given buffer.

 

Quality control & Technical tips

 


Pf1 price list 2017 (in Euro, )

  Item Id. Amount in mg Price, (w/o shipping costs)
Pf1 cosolvent, protease free (in 10mM K-phosphate buffer pH7.6, 2mM MgCl2 and 0.05% NaN3)
P-50-P
50
399.00
P-100-P
100
599.00
P-300-P
300
1369.00
P-500-P
500
2129.00
   
 
   
 
Pf1 cosolvent, RNAase and protease free (in 10mM K-phosphate buffer pH7.6, DEPC-treated water)
P-50-RNA
50
439.00
P-100-RNA
100
659.00
P-300-RNA
300
1489.00
P-500-RNA
500
2309.00
     
Custom buffer

including unlabeled isotope salts

50
+149.00
including deuterium oxide or other labeled compounds  
please enquire
     
     
     
     

 


References

  • References from Art Pardi, Marius Clore, Ad Bax, Gottfried Otting, and other World NMR Experts
  • Papers that refer to pf1 magnetic resonance co-solvent from ASLA Biotech.

 


Our Customers Worldwide

+371-67428426

Ratsupites 1, k1. Riga, LV-1067, Latvia