Dendrimers as protein denaturants

Dendrimers are well-defined chemical compounds with a characteristic branching pattern that
gives rise to attractive features such as antibacterial and antitumor activities as well as drug
delivery properties.  In addition, dendrimers can solubilize prion protein lumps at very low
concentrations, but their mode of action is unclear. Poly(propylene imine) dendrimers based on
di-aminobutane (DAB) and alter with guanidinium surface groups reduce insulin thermostability
and solubility considerably at microgram per microliter concentrations, while urea alter groups
have hardly any effect. Proteins can interact with both dendrimer surface and interior. The pH-
dependence release that interconnections are mainly mediated by electrostatics, confirmed by
studies on four other proteins.  Ability to precipitate and disrupt are positively correspond, in
contrast to conventional small-molecule denaturants and stabilizers, indicating that surface
immobilization of denaturing groups profoundly affects its interactions with proteins. (1)
Stabilizing effect of small concentrations of PAMAM dendrimers at the insulin aggregation
Dendrimers action on proteins and peptides has a double and disputatable character. On one
hand, they dissolve prion protein and amyloid fibrils lumps, which are otherwise only soluble in
solvents containing both detergents and high denaturant concentrations.  On the other hand they
are able to destabilize proteins in generation dependent manner.In present work we estimated the
influence of small concentrations (up to 1.4 μg/ml) of cationic, neutral and anionic
poly(amidoamine) dendrimers of 3rd and 4th generations on dithiotreitol induced aggregation of
insulin. It was found that cationic dendrimers decreased the insulin aggregation, while anionic
and neutral ones did not. At the same time, destabilizing effect of dendrimers on insulin structure
was not observed. The conclusion was made that small concentrations of dendrimers can be
applied to prevent or decrease the formation of misfolded structures of protein. (2)

Small concentrations of dendrimers did not disturb structure of insulin.Cationic dendrimers

decreased aggregation of insulin.Anionic dendrimers had no effect on secondary structure of
insulin.
Interactions between PAMAM dendrimers and bovine serum albumin
Dendrimers are a new class of polymeric materials. They are spherical, highly branched,
monodisperse macromolecules. Due to their structure, dendrimers promise to be new, effective
biomedical materials as oligonucleotide transfection agents and drug carriers. More information
about biological properties of dendrimers is critical for further investigation of dendrimers in
therapeutic applications.The mechanism of interactions between polyamidoamine (PAMAM)
dendrimers and bovine serum albumin (BSA) was examined. PAMAM dendrimers are based on
an ethylenediamine core and branched units are constructed from both methyl acrylate and
ethylenediamine. We used three types of PAMAM dendrimers with different surface groups (–
COOH, –NH2, –OH). As BSA contains two tryptophan residues we were able to evaluate
dendrimers influence on protein molecular conformation by measuring the changes in the
fluorescence of BSA in the presence of dendrimers. Additionally experiments with a fluorescent
probe 1-anilinonaphthalene-8-sulfonic acid (ANS) were carried out. The differential scanning
calorimetry (DSC) was chosen to investigate impact on protein thermal stability upon the
dendrimers. (3)

Effects of PAMAM Dendrimer Salt Solutions on Protein Stability

Dendrimers are widely used for biological applications. However, their effect on protein stability
has not been studied substantially. Typically, charged cationic dendrimers such as PAMAM
dendrimers tend to disrupt proteins due to the cooperative binding of surface groups to the
protein surface. We have studied the effect of PAMAM dendrimer salt solutions on protein
stability both experimentally and arithmatically. We show that the effect of dendrimers on
protein stability depends on the choice of counterion (e.g., dihydrogen phosphate salts reduce the
rate of protein aggregation, while chloride salts increase the rate of protein aggregation). In the
presence of dihydrogen phosphate and sulfate counterions, the binding of dendrimers to the
protein surface is limited (when compared to chloride and thiocyanate), which increases the
conformational stability of proteins.This is the first study that has shown that PAMAM
dendrimer salts can significantly suppress the aggregation of proteins. (4)

References
1. Giehm, L., Christensen, C., Boas, U., Heegaard, P. M., & Otzen, D. E. (2008). Dendrimers
destabilize proteins in a generation‐dependent manner involving electrostatic
interactions. Biopolymers: Original Research on Biomolecules, 89(6), 522-529.
2. Nowacka, O., Shcharbin, D., Klajnert-Maculewicz, B., & Bryszewska, M. (2014). Stabilizing
effect of small concentrations of PAMAM dendrimers at the insulin aggregation. Colloids and
Surfaces B: Biointerfaces, 116, 757-760.
3. Klajnert, B., Stanisławska, L., Bryszewska, M., & Pałecz, B. (2003). Interactions between
PAMAM dendrimers and bovine serum albumin. Biochimica et Biophysica Acta (BBA)-Proteins
and Proteomics, 1648(1-2), 115-126.
4. Shukla, D., Schneider, C. P., & Trout, B. L. (2011). Effects of PAMAM dendrimer salt
solutions on protein stability. The Journal of Physical Chemistry Letters, 2(14), 1782-1788.