Nucleation of protein fibrillation by nanoparticles

Linse et al. 10.1073/pnas.0701250104.

Supporting Information

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SI Table 1
SI Text
SI Figure 7

SI Figure 7

Fig. 7. Strategy for the PCR production and cloning of a synthetic gene coding for the human b₂m protein. (Upper) Schematic representation of the clone production. The synthetic gene, with codons optimized for expression in E. coli, was constructed from four overlapping oligonucleotides (a, b, c, and d) and two end primers (start and stop) by using PCR and cutting with restriction enzymes NdeI and SacI. The intervening purification steps using agarose gel electrophoresis and spin columns are not shown. The synthetic gene was cloned into the PetSac vector (modified Pet3a vector with NdeI and SacI cloning sites). (Lower) The nucleotide sequences of the four overlapping oligonucleotides (a, b, c, and d) and the two end primers (start and stop).

Table 1. Solution conditions of the fibrillation experiments in the presence and absence of copolymer nanoparticles

Sodium phosphate, mM	NaCl, mM	b₂m, mM	No. of samples
10	None added	40	120
6*	50	125	120
6*	50	40	96
6*	50	40	48
20*	50	40	48
20*	50	40	120
20*	50	40	90
20*	40	40	90

An additional group of 90 samples were studied without or with quantum dots, carbon nanotubes, or cerium oxide particles. All solutions were at pH 2.5, 37°C, with 0.02% NaN₃.

^*These samples contained 3 mM Tris/HCl from the gel filtration buffer.