The Limitations of an Exclusively Colloidal View of Protein Solution Hydrodynamics and Rheology

Proteins are complex macromolecules with dynamic conformations. They are charged like colloids, but unlike colloids, charge is heterogeneously distributed on their surfaces. Here we overturn entrenched doctrine that uncritically treats bovine serum albumin (BSA) as a colloidal hard sphere by elucidating the complex pH and surface hydration-dependence of solution viscosity. We measure the infinite shear viscosity of buffered BSA solutions in a parameter space chosen to tune competing long-range repulsions and short-range attractions (2 mg/mL ≤ [BSA] ≤ 500 mg/mL and 3.0 ≤ pH ≤ 7.4). We account for surface hydration through partial specific volume to define volume fraction and determine that the pH-dependent BSA intrinsic viscosity never equals the classical hard sphere result (2.5). We attempt to fit our data to the colloidal rheology models of Russel, Saville, and Schowalter (RSS) and Krieger-Dougherty (KD), which are each routinely and successfully applied to uniformly charged suspensions and to hard-sphere suspensions, respectively. We discover that the RSS model accurately describes our data at pH 3.0, 4.0, and 5.0, but fails at pH 6.0 and 7.4, due to steeply rising solution viscosity at high concentration. When we implement the KD model with the maximum packing volume fraction as the sole floating parameter while holding the intrinsic viscosity constant, we conclude that the model only succeeds at pH 6.0 and 7.4. These findings lead us to define a minimal framework for models of crowded protein solution viscosity wherein critical protein-specific attributes (namely, conformation, surface hydration, and surface charge distribution) are addressed.     

DONNAN EQUILIBRIUM AND THE PHYSICAL PROPERTIES OF PROTEINS : III. VISCOSITY.

1. Gelatin solutions have a high viscosity which in the case of freshly prepared solutions varies under the influence of the hydrogen ion concentration in a similar way as the swelling, the osmotic pressure, and the electromotive forces. Solutions of crystalline egg albumin have under the same conditions a comparatively low viscosity which is practically independent of the pH (above 1.0). This difference in the viscosities of solutions of the two proteins seems to be connected with the fact that solutions of gelatin have a tendency to set to a Jelly while solutions of crystalline egg albumin show no such tendency at low temperature and pH above 1.0. 2. The formulæ for viscosity demand that the difference in the order of magnitude of the viscosity of the two proteins should correspond to a difference in the relative volume occupied by equal masses of the two proteins in the same volume of solution. It is generally assumed that these variations of volume of dissolved proteins are due to the hydration of the isolated protein ions, but if this view were correct the influence of pH on viscosity should be the same in the case of solutions of gelatin, of amino-acids, and of crystalline egg albumin, which, however, is not true. 3. Suspensions of powdered gelatin in water were prepared and it was found, first, that the viscosity of these suspensions is a little higher than that of gelatin solutions of the same concentration, second, that the pH influences the viscosity of these suspensions similarly as the viscosity of freshly prepared gelatin solutions, and third, that the volume occupied by the gelatin in the suspension varies similarly as the viscosity which agrees with the theories of viscosity. It is shown that this influence of the pH on the volume occupied by the gelatin granules in suspension is due to the existence of a Donnan equilibrium between the granules and the surrounding solution.     

Microwave-aided binding of gold-protein-ligand (GPL) complexes. Light microscopic observations in the rat brain.

We describe a rapid method for the preparation and binding site labeling of cryostat sections for use in light microscopy. Instead of using antibodies to bind to specific sites, substance P, delta-sleep-inducing peptide, oxytocin, and dopamine were covalently attached to BSA and then the BSA-ligand complex was adsorbed on 5-nm colloidal gold particles. Bioassays carried out on isolated organs indicated that the physiological activity of the ligand GPL complex was maintained. Most of the technical steps included use of an ordinary microwave oven (MWO), with tissues exposed for less than 1 min in any given step. Cryostat sections of unfixed rat brain were pre-incubated for 50 sec in the MWO in a Tris-buffered solution (pH 7.4) containing 1.5% BSA, then further incubated for 50 sec in the MWO in Tris-buffered solution containing 1% gelatin and the diluted colloidal gold suspension. After washing, the preparations were postfixed for 30 sec in the MWO in 5% formaldehyde solution, pH 7.4. Finally, the cell-bound gold particles were enlarged by a silver-enhancing process and counterstained. Preparations observed at high magnification provided excellent resolution of the cell binding sites. Positive and negative controls performed by addition of BSA-conjugated ligands to the pre-incubation and incubation medium, and displacement of the markers by an excess of unbound ligand in the pre-incubation or the incubation medium, showed the specificity of the tissue labeling.     

5-Fluorouracil-Loaded BSA Nanoparticles: Formulation Optimization and In Vitro Release Study

Over the past few decades, there has been considerable interest in developing protein nanoparticles as drug delivery devices. The underlying rationale is their exceptional characteristics, namely biodegradability and nonantigenicity. Herein, phase separation method was used to prepare 5-fluorouracil-loaded bovine serum albumin (BSA) nanoparticles. Drug release was tracked by continuous flow dialysis technique. Effect of process variables on loading efficiency of 5-fluorouracil was investigated and optimized through Taguchi’s M16 design with the amount of entrapped drug as response. Optimum condition was found to be 2 mg/mL of 5-fluorouracil, 3.7 mL of added ethanol, 176 μL of glutaraldehyde, drug–protein incubation time of 30 min, and pH of 8.4 for 200 mg of BSA in 2 mL drug solution. pH had the most noticeable effect on the amount of entrapped drug, but glutaraldehyde had the least. Mean diameter and zeta potential of fabricated nanoparticles under these conditions were 210 nm and −31.7 mV, respectively. Drug-loaded BSA nanoparticles suspension maintained constant release of drug for 20 h under experimental conditions, so this colloidal drug carrier is capable of releasing drug in a sustained manner.     

Low molecular weight chitosan-g-l-phenylalanine: Preparation,characterization, and complex formation with DNA

The grafting of l-phenylalanine onto low molecular weight chitosan is accomplished by using carbodiimide as a coupling agent. As increase in the amount of phenylalanine in feed, the grafting chain length increases, while a number of grafting chains hardly change. The obtained product, LMWCts-g-Phe, performs sphere with an average size of ～80 nm when the % grafting is less than 123. The complexes of the LMWCts-g-Phe and DNA (LMWCts-g-Phe/DNA) prepared by a complex coacervation method possess various shapes with an average size of ～50–150 nm and a negatively charged surface. The LMWCts-g-Phe and its complex show very reduced toxicity to fibroblast cells. The release of DNA from the complex is very fast in high pH media (tris buffer, pH 8.0 and carbonate buffer, pH 9.5), and relatively slow or more sustainable in neutral and low pH ones (PBS, pH 7.4 and citric acid/trisodium citrate buffer, pH 3.0). The results suggest that the LMWCts-g-Phe be an alternative promising carrier for negatively charged active molecules.     

Controlling protein transport in ultrafiltration using small charged ligands

Previous studies have demonstrated that protein transport during ultrafiltration can be strongly influenced by solution pH and ionic strength. The objective of this study was to examine the possibility of controlling protein transmission using a small, highly charged ligand that selectively binds to the protein of interest. Experiments were performed using bovine serum albumin and the dye Cibacron Blue. Protein sieving data were obtained with essentially neutral and negatively charged versions of a composite regenerated cellulose membrane to examine the effects of electrostatic interactions. The addition of only 1 g/L of Cibacron Blue to an 8 g/L BSA solution reduced the BSA sieving coefficient through the negatively-charged membrane by more than two orders of magnitude, with this effect being largely eliminated at high salt and with the neutral membrane. Protein sieving data were in good agreement with model calculations based on the partitioning of a charged sphere in a charged pore accounting for the change in net protein charge due to ligand binding and the increase in solution ionic strength due to the free ligand in solution.     

Adsorption of BSA on strongly basic chitosan: Equilibria

Equilibrium isotherms for adsorption of bovine serum albumin (BSA) on a new adsorbent, a strongly basic crosslinked chitosan (Chitopearl 2503), which is hard and is not compressed by pressure in a column, have been presented and compared with diethylaminoethyl (DEAE) Sepharose Fast Flow (hard gel). In Chitopearl 2503, when only buffer existed in the BSA solution, the isotherm was not affected by the initial concentration of BSA but it was affected by pH considerably. The isotherm was favorable when pH >/= pl ( congruent with 4.8). When NaCl existed in the BSA solution, the amount of BSA absorbed on the resin decreased with increasing concentration of NaCl. When the concentration of NaCl was 200 mol/m(3), the resin did not adsorb BSA at all. The equilibrium data were correlated by the Langmuir equation reasonably well. The BSA may be adsorbed mainly by electrostatic attraction between negatively charged BSA and positively charged quanternary ammonium groups at pH > pl and by protonation reaction of the primary ammonium groups by weak acid groups of BSA at pH = pl. These are confirmed by measuring the amount of inorganic ion exchanged for BSA. In DEAE Sepharose Fast Flow, the isotherm was favorable when pH > pl but unfavorable ar pH = pl. The saturation capacity of BSA on Chitopearl 2503 is about 1.3 to 2.2 times larger than that on DEAE Sepharose Fast Flow. (c) 1994 John Wiley & Sons, Inc.     

Expression,purification, and refolding of a recombinant human bone morphogenetic protein 2 in vitro

In this work, the recombinant human bone morphogenetic protein 2 (rhBMP-2) gene was cloned from MG-63 cells by RT-PCR, and the protein was expressed in Escherichia coli expression system, purified by Ni–NTA column under denaturing conditions and refolded at 4 °C by urea gradient dialysis. We found that the protein refolding yield was increased with the increase of pH value from pH 6.0 to pH 9.0. The yield was 42% and 96% at pH 7.4 and pH 9.0, respectively, while that at pH 6.0 was only 3.4%. The cell culture results showed that the rhBMP-2 refolded at pH 7.4 urea gradient dialysis had higher biological activity for MG-63 cell proliferation and differentiation than that refolded at pH 9.0 since pH 7.4 is closer to the conditions in vivo leading to the formation of dimers through the interchain disulfide bond. Moreover, the biological activity for MG-63 was promoted with the increase of rhBMP-2 concentration in the cell culture medium. This work may be important for the in vitro production and biomedical application of rhBMP-2 protein.     

pH and cell volume effects on H2O and phosphoryl resonance splitting in rapid-spinning NMR of red cells

Two resonances are seen in the ¹H-NMR spectrum of water in erythrocyte suspensions spun at the magic angle, a broad signal from water inside the cells and a sharp signal from extracellular water. The splitting is a result of a true chemical shift difference between the two populations, as bulk magnetic susceptibility effects are negated at the magic angle. The pH dependence of this chemical shift difference in erythrocyte suspensions was investigated. Splittings of 16.7 ± 0.1, 18.9 ± 0.9, and 21.0 ± 0.2 Hz were observed at pH 6.0, 7.0, and 8.5, respectively; however, this was accompanied by a change in the mean cell volume. To account for any contribution from the volume change, the osmolality of the pH 6.0 and 8.5 suspensions was adjusted to equalize the cell volume between samples at the three pHs. Under these conditions, the splitting was 18.3 ± 0.1 and 18.6 ± 0.1 Hz at pH 6.0 and 8.5, respectively. Thus the observed chemical shift difference between the two water resonances was independent of pH. Therefore the splitting of the water resonance was concluded to be directly proportional to the protein concentration within the cell. Measurements of the magnetic susceptibility difference between the two compartments were also carried out, yielding a value of 2.0 ± 0.2 × 10⁻⁷ (SI units) for erythrocytes in isotonic saline at pH 7.0.     

Osmotic pressure and aggregate shape in BSA/poly(ethylene glycol)-lipid/Dextran solutions

In this work we study the colloidal osmotic pressure (COP) and aggregate shape in phosphate saline buffer solutions (pH 7.4) containing bovine serum albumin (BSA), poly(ethylene glycol) lipid (PEG(2000)-PE) and Dextran (Dx). Dx was added to the BSA/PEG(2000)-PE system in order to increase the COP of the solution to levels comparable to the COP of healthy adults, with the aim of using the solution as a blood COP regulator. Dynamic light scattering and small angle X-ray scattering results shown the formation of BSA/PEG(2000)-PE/Dx aggregates in the solution. Osmometry results shown that the addition of Dx to the BSA/PEG(2000)-PE system could successfully increase the COP, through the formation of BSA/PEG(2000)-PE/Dx aggregates. The BSA/PEG(2000)-PE/Dx solutions attained COP=15 mm Hg, representing 60% of COP measured for healthy adults.     

Hydrolysis of Chitosan Sulfate with an Enzyme Complex from Streptomyces kurssanovii

The possibility of enzymatic hydrolysis of chitosan was shown. The optimum conditions for the process are sodium acetate buffer pH 6.0, 37°C, 24 h, and a chitosan sulfate–protein volume ratio of 500 : 1 in the enzyme preparation. During hydrolysis, the intrinsic viscosity of chitosan sulfate solution decreased by a factor of 2.7.     

DONNAN EQUILIBRIUM AND THE PHYSICAL PROPERTIES OF PROTEINS : IV. VISCOSITY—Continued.

1. The proof is completed that the influence of electrolytes on the viscosity of suspensions of powdered particles of gelatin in water is similar to the influence of electrolytes on the viscosity of solutions of gelatin in water. 2. It has been suggested that the high viscosity of proteins is due to the existence of a different type of viscosity from that existing in crystalloids. It is shown that such an assumption is unnecessary and that the high viscosity of solutions of isoelectric gelatin can be accounted for quantitatively on the assumption that the relative volume of the gelatin in solution is comparatively high. 3. Since isoelectric gelatin is not ionized, the large volume cannot be due to a hydration of gelatin ions. It is suggested that this high volume of gelatin solutions is caused by the existence in the gelatin solution of submicroscopic pieces of solid gelatin occluding water, the relative quantity of which is regulated by the Donnan equilibrium. This would also explain why the influence of electrolytes on the viscosity of gelatin solutions is similar to the influence of electrolytes on the viscosity of suspensions of particles of gelatin. 4. This idea is supported by experiments on solutions and suspensions of casein chloride in which it is shown that their viscosity is chiefly due to the swelling of solid particles of casein, occluding quantities of water regulated by the Donnan equilibrium; and that the breaking up of these solid particles into smaller particles, no longer capable of swelling, diminishes the viscosity. 5. This leads to the idea that proteins form true solutions in water which in certain cases, however, contain, side by side with isolated ions and molecules, submicroscopic solid particles capable of occluding water whereby the relative volume and the viscosity of the solution is considerably increased. This accounts not only for the high order of magnitude of the viscosity of such protein solutions but also for the fact that the viscosity is influenced by electrolytes in a similar way as is the swelling of protein particles. 6. We therefore reach the conclusion that there are two sources for the viscosity of protein solutions; one due to the isolated protein ions and molecules, and the other to the submicroscopic solid particles contained in the solution. The viscosity due to the isolated molecules and ions of proteins we will call the general viscosity since it is of a similar low order of magnitude as that of crystalloids in solution; while the high viscosity due to the submicroscopic solid protein particles capable of occluding water and of swelling we will call the special viscosity of protein solutions. Under ordinary conditions of hydrogen ion concentration and temperature (and in not too high a concentration of the protein in solution) the general viscosity due to isolated ions and molecules prevails in solutions of crystalline egg albumin and in solutions of metal caseinates (where the metal is monovalent) while under the same conditions the second type of viscosity prevails in solutions of gelatin and in solutions of acid-salts of casein; and also in solutions of crystalline egg albumin at a pH below 1.0 and at higher temperatures. The special viscosity is higher in solutions of gelatin than of casein salts for the probable reason that the amount of water occluded by the submicroscopic solid gel particles in a gelatin solution is, as a rule, considerably higher than that occluded by the corresponding particles of casein.     

Influence of Colloidal Iron on the Respiration of a Species of the Genus Acinetobacter

Washed suspensions of Acinetobacter sp. isolated from water caused the precipitation of iron from a suspension of colloidal ferric iron at pH 6.0 and 7.6. Iron-encrusted cells of the bacterium formed large aggregates. The amount of iron removed from the colloidal preparation in the form of aggregates was from 21 to 52% at pH 7.6 and 49% an pH 6.0 by the bacterial cells. Endogenous respiration rates of the iron-encrusted cells were from 32 to 72% lower than the rates for unencrusted cells. Respiration rates, measured polarographically in the presence of glucose, were also greatly reduced by the coating of iron on the cells.     

Temperature dependence of macromolecular interactions in dilute and concentrated hemoglobin solutions

We studied the temperature-dependent effects of intramolecular interactions on the mutual diffusion coefficient of normal human oxygenated hemoglobin in salt solution. We used photon correlation spectroscopy to observe this temperature dependence of the mutual diffusion coefficient of two protein concentrations (1.25 and 17.0 g %) between 13 and 37°C. This coefficient was our probe for monitoring temperature-dependent structural changes of hydrated hemoglobin in solution. Comparison of our measured diffusion coefficient with that predicted by the Stokes-Einstein relationship in terms of solvent or solution viscosity showed a clear transition in the conformation of hemoglobin at approximately 22°C, independent of the hemoglobin concentration. We postulated that at this physiological temperature, a considerable quaternary rearrangement of the hemoglobin chains takes place. We believe this rearrangement changes the effective volume and the hydration sphere of the hemoglobin macromolecule.     

Effect of pH on protein distribution in electrospun PVA/BSA composite nanofibers

We examine the protein distribution within an electrospun polymer nanofiber using polyvinyl alcohol and bovine serum albumin as a model system. We hypothesize that the location of the protein within the nanofiber can be controlled by carefully selecting the pH and the applied polarity of the electric field as the pH affects the net charge on the proteins. Using fluorescently labeled BSA and surface analysis, we observe that the distribution of BSA is affected by the pH of the electrospinning solution. Therefore, we further probe the relevant forces on the protein in solution during electrospinning. The role of hydrodynamic friction was assessed using glutaraldehyde and HCl as a tool to modify the viscosity of the solution during electrospinning. By varying the pH and the polarity of the applied electric field, we evaluated the effects of electrostatic forces and dielectrophoresis on the protein during fiber formation. We surmise that electrostatic forces and hydrodynamic friction are insignificant relative to dielectrophoretic forces; therefore, it is possible to separate species in a blend using polarizability contrast. A coaxial distribution of protein in the core can be obtained by electrospinning at the isoelectric point of the protein, whereas surface enrichment can be obtained when the protein carries a net charge.     

Water-soluble complexes through coulombic interactions between bovine serum albumin and anionic polyelectrolytes grafted with hydrophilic nonionic side chains

The interaction between bovine serum albumin (BSA) and the anionic graft copolymers poly(sodium acrylate-co-sodium 2-acrylamido-2-methyl-1-propanesulfonate)-graft-poly(N,N-dimethylacrylamide) (P(NaA-co-NaAMPS)-g-PDMAMx) was investigated within the acid pH region, 2 < or = pH < or = 7. The weight percentage, x, of the poly(N,N-dimethylacrylamide) (PDMAM) side chains varied from 0 up to 75% (w:w). When BSA and P(NaA-co-NaAMPS)-g-PDMAMx are oppositely charged, i.e., when pH is lower than the isoelectric point of BSA, the two macromolecules associate through Coulombic attractions. When the anionic graft copolymer is rich enough to the nonionic PDMAM side chains, x > or = 50% w:w, the associative phase separation is practically prevented, as revealed by the turbidimetric study of the BSA/P(NaA-co-NaAMPS)-g-PDMAMx mixtures in aqueous solution vs pH. In addition, the viscosity measurements support the formation through a charge neutralization process of a rather compact protein-polyelectrolyte complex stabilized by the hydrophilic PDMAM side chains grafted onto the anionic copolymer backbone.     

Sequential adsorption of F(ab')(2) and BSA on negatively and positively charged polystyrene latexes

The aim of the present work is to study the sequential adsorption of F(ab')(2) and bovine serum albumin (BSA) molecules adsorbed onto positively and negatively charged polystyrene latexes. Cationic and anionic latexes were prepared by emulsifier-free emulsion polymerization. Adsorptions of F(ab')(2) on both latexes at a low ionic strength and different pHs were performed. The cationic latex showed a higher adsorption of F (ab')(2) molecules over a range of pH, which could be due to the formation of multilayers. Sequential adsorption of anti-CRP F(ab')(2) and monomeric BSA were performed at two different pre-adsorbed F(ab')(2) amounts on both types of latex. Displacement of F(ab')(2) occurred only when the preadsorbed amounts were larger than a certain critical value, which depends on the adsorption pH. A greater displacement of larger preadsorbed amounts might be the result of a weaker contact between the protein molecules and the polystyrene surface. The displacement of F(ab')(2) previously adsorbed onto both latexes occurred due to pH changes, an increase of ionic strength and the presence of BSA molecules. The effect caused by these three factors was studied independently. The main factors in the desorption of F(ab')(2) on the anionic latex are the changes in pH and ionic strength, whereas on the cationic latex the desorption is mainly caused by the increase of the ionic strength and the presence of BSA. The colloidal stability of the immunotatex was improved by BSA adsorption, especially on cationic latex. (c) 1995 John Wiley & Sons, Inc.     

Rapid evolution of arginine deiminase for improved anti-tumor activity

Arginine deiminase (ADI), an arginine-degrading enzyme, has been studied as a potential anti-cancer agent for inhibiting arginine-auxotrophic tumors, such as melanomas and hepatocellular carcinomas. Based on our preliminary results, it was noticed that the optimum pH of ADI from Pseudomonas plecoglossicida (PpADI) was 6.0, and less than 10% of the activity was retained at pH 7.4 (pH of human plasma). Additionally, the K _m value for wild-type ADI (WT-ADI) was 2.88 mM (pH 6.0), which is over 20 times of the serum arginine level (100–120 μM). These are two major limitations for PpADI as a potential anti-cancer drug. A highly sensitive and efficient high-throughput screening strategy based on a modified diacetylmonoxime–thiosemicarbazide method was established to isolate ADI mutants with higher activity and lower K _m under physiological pH. Three improved mutants was selected from 650 variants after one round of ep-PCR, among which mutant 314 (M314: A128T, H404R, I410L) exhibiting the highest activity. Interestingly, sequence alignment shows that three amino acid substitutes in M314 are coincident with corresponding residues in ADI from Mycoplasma arginini. The specific activity of M314 (9.02 U/mg) is over 20-fold higher than that of WT-ADI (0.44 U/mg) at pH 7.4, and the K _m value was reduced to 0.65 mM (pH 7.4). Noticeably, the pH optimum was shifted from 6.0 to 6.5 in M314. Homology model of M314 was constructed to understand the molecular basis of the improved enzymatic properties. This work could provide promising drug candidate for curing arginine-auxotrophic cancers.     

银胶中三螺旋RNA poly（rU）．poly（rA）．poly（rU）的付立叶表面增强拉曼光谱研究

采用近红外付立叶拉曼光谱研究了三螺旋ＲＮＡ（ｒＵ）．ｐｏｌｙ（ｒＡ）．ｐｏｌｙ（ｒＵ）在溶液中的构象和在银胶中的表面增强拉曼散行为。结果表明在溶液中,该三螺旋ＲＮＡ分子中以Ｗａｔｓｏｎ－Ｃｒｉｃｋ碱基酸对的两条链处于Ａ－构型,而第二条嘧啶链处于Ｃ２’－ｅｎｄｏ／ａｎｔｉ构象。在银胶中,该三螺旋ＲＮＡ的表面增强拦曼效应明显。与溶液状态下相比,８３５和８１９ｃｍ＾－１谱带的出现暗示该三螺旋ＲＮＡ吸附到     

Viscous cosolvent effect on the ultrasonic absorption of bovine serum albumin.

Protein-ligand binding and enzyme activity have been shown to be regulated by solvent viscosity, induced by the addition of viscous cosolvents. This was indirectly interpreted as an effect on protein dynamics. However, viscous cosolvents might affect dynamic, e.g., viscosity, as well as thermodynamic properties of the solution, e.g., activity of solution components. This work was undertaken to examine the effect of viscous cosolvent on the structural dynamics of proteins and its correlation with dynamic and thermodynamic solution properties. For this purpose we studied the effect of viscous cosolvent on the specific ultrasonic absorption, delta mu, of bovine serum albumin, at pH = 7.0 and at 21 degrees C, and frequency range of 3-4 MHz. Ultrasonic absorption (UA) directly probes protein dynamics related to energy dissipation processes. It was found that the addition of sucrose, glycerol, or ethylene glycol increased the BSA delta mu. This increase correlates well with the solvent viscosity, but not with the cosolvent mass concentration, activity of the solvent components, dielectric constant, or the hydration of charged groups. On the grounds of these results and previously reported findings, as well as theoretical considerations, we propose the following mechanism for the solvent viscosity effect on the protein structural fluctuations, reflected in the UA: increased solvent viscosity alters the frequency spectrum of the polypeptide chain movements; attenuating the fast (small amplitude) movements, and enhancing the slow (large amplitude) ones. This modulates the interaction strength between the polypeptide and water species that "lubricates" the chain's movements, leading to larger protein-volume fluctuation and higher ultrasonic absorption. This study demonstrates that solvent viscosity is a regulator of protein structural fluctuations.