Reviewing the thermodynamic parameters characterizing self-association and ligand binding of proteins at 25 OC, we find AGO, AHo, AS\", and ACpo are often all of negative sign. It is thus not possible to account for the stability of association complexes of proteins on the basis of hydrophobic interactions alone. We present a conceptual model of protein association consisting of two steps: the mutual penetration of hydration layers causing disordering of the solvent followed by further short-range interactions. The net AGO for the complete association process is primarily determined by the positive entropy change accompanying the first step and the negative enthalpy change of the second step. On the basis of the thermochemical behavior of small molecule interactions, we conclude that the strengthening of hydrogen bonds in the I n the past decade, a complete thermodynamic description of the self-association of many proteins and their interactions From the Laboratory of Molecular Biology (P.D.R.) and Laboratory of Nutrition and Endocrinology (S.S.), National Institute of Arthritis, Metabolism and Digestive Diseases, National Institutes of Health, Bethesda, Maryland 20205. Received September 23, 1980. low dielectric macromolecular interior and van der Waals' interactions introduced as a consequence of the hydrophobic effect are the most important factors contributing to the observed negative values of AHo and ASo and hence to the stability of protein association complexes. The X-ray crystallographic structures of these complexes are consonant with this analysis. The tendency for protein association reactions to become entropy dominated and/or entropy-enthalpy assisted at low temperatures and enthalpy dominated at high temperatures (a consequence of the typically negative values of AC,\") arises from the diminution of the hydrophobic effect with increasing temperature which is a general property of the solvent, water. with small molecular substrates has become available. Concomitantly, X-ray crystallography has provided a detailed picture of some of these associations, and this has stimulated a number of theoretical studies (Levitt & Warshel, 1975; Gelin & Karplus, 1975; Chothia & Janin, 1975), based upon energetic considerations, to account for these structures. The This article not subject to U S . Copyright. Published 1981 by the American Chemical Society T H E R M O D Y N A M I C S O F P R O T E I N A S S O C I A T I O N V O L . 2 0 , N O . 1 1 , 1 9 8 1 3097 Table I: Thermodynamics of Protein Association' association process A G \" ~ AiY A s o , A c p o (kcal mol-') (kcal mol-l) (cal K-I mol-') (cal K-I mol-I) refb trypsin (bovine) + inhibitor (soybean) -14.6 8.6 78 -440 c, d deoxyhemoglobin S gelation -3.4 2.0 18 -200 e, f lysozyme self-association (indefinite) -3.9 -6 .4 -8.3 g glucagon trimerization -12.1 -3 1 -64 -430 h, i hemoglobin t haptoglobin -11.5 -3 3 -7 3 -940 i a-chymotrypsin dimerization -7.1 -35 -9 5 k, I S-peptide + S-protein (ribonuclease) -13 -40 -90 -1100 m, n All thermodynamic parameters expressed per mole of complex formed except the indefinite association cases of hemoglobin S and lysozyme for which the mole refers to the monomeric protein reacted. Unitary entropy and free energy are given for processes of defined stoichiometry. Standard states are hypothetical 1 M protein, pH at which the reaction was measured. All pHs were close to 7 except for trypsin, pH 5, haptoglobin, pH 5.5, and glucagon, pH 10.5. All data for 25 \"C except glucagon, T = 30 \"C. ence is to calorimetric work and the second is to X-ray crystallographic structure determination. al. (1974). e Rosset al. (1977). Wishner e t al. (1975). g Banerjee et al. (1975). Johnson et al. (1979). * Sasaki et al. (1975). For each entry, the first referSweet et Baugh & Trowbridge (1972). Lavialle et al. (1974). Shiao & Sturtevant (1969). lVandlen &Tulinsky (1973). Hearn et al. (1971). Wyckoff e t al. (1970). methodology and problems involved in such calculations have been critically reviewed by NBmethy & Scheraga (1977). In this paper we review the thermodynamics of protein association processes for the examples best characterized in terms of their chemistry and structure. From this survey we find that the thermodynamic parameters AGO, Ai?, AS\", and ACpo are predominantly of negative sign. This result poses severe difficulties for interpretations of protein association based upon the entropically driven hydrophobic effect. The aim of this paper is to attempt to account for the signs and magnitudes of these thermodynamic parameters for protein association reactions in terms of known molecular forces and the thermochemistry of small molecule interactions.

Thermodynamics of protein association reactions: forces contributing to stability

Nitric oxide (NO.), a potentially toxic molecule, has been implicated in a wide range of biological functions. Details of its biochemistry, however, remain poorly understood. The broader chemistry of nitrogen monoxide (NO) involves a redox array of species with distinctive properties and reactivities: NO+ (nitrosonium), NO., and NO- (nitroxyl anion). The integration of this chemistry with current perspectives of NO biology illuminates many aspects of NO biochemistry, including the enzymatic mechanism of synthesis, the mode of transport and targeting in biological systems, the means by which its toxicity is mitigated, and the function-regulating interaction with target proteins.

https://science.sciencemag.org/content/sci/258/5090/1898.full.pdf

Biochemistry of nitric oxide and its redox-activated forms

A geometric approach to macromolecule-ligand interactions

Novel crosslinking methods to design hydrogels

Publisher Summary This chapter focuses on structure and expression of interleukin-6 (IL-6), which is a cytokine with pleiotropic activities that plays a central role in host defense. IL-6 can exert growth-inducing, growth-inhibitory, and differentiation-inducing activities, depending on the target cells. These activities include (1) terminal differentiation (secretion of immunoglobulins) in B cells and (2) growth promotion on various B cells. IL-6 has been implicated in the pathology of many diseases including multiple myeloma, mesangial proliferative glomerulonephritis, rheumatoid arthritis, and acquired immunodeficiency syndrome (AIDS). Selective inhibition of the synthesis or of the action of IL-6 may have therapeutic benefit against the IL-6-associated diseases. On the other hand, IL-6 has potent antitumor activity against certain types of tumors. Application of IL-6 is promising in cancer treatment as well as in treatment of radiation- or chemotherapy-induced myelosuppression. The cell biology of the intracellular events that link transduction to gene regulation is an important area, and work on these topics helps to understand such phenomena as multifunction of IL-6 and bidirectional effects of cell growth depending on the cell type.

Interleukin-6 in biology and medicine

Free radicals have been postulated to play an important role as mediators in the pathogenesis of shock syndrome and multiple-organ failure. We attempted to directly detect the increased formation of radicals by Electron Spin Resonance (ESR) in animal models of shock, namely the endotoxin (ETX) shock or the hemorrhagic shock of the rat. In freeze-clamped lung tissue, a small but significant increase of a free radical signal was detected after ETX application. In the blood of rats under ETX shock, a significant ESR signal with a triplet hyperfine structure was observed. The latter ESR signal evolved within several hours after the application of ETX and was localized in the red blood cells. This signal was assigned to a nitric oxide (NO) adduct of hemoglobin with the tentative structur ((a2+ NO)/23+)2. The amount of hemoglobin-NO formed, up to 0.8% of total hemoglobin, indicated that under ETX shock a considerable amount of NO was produced in the vascular system. This NO production was strongly inhibited by ...

Formation of free radicals and nitric oxide derivative of hemoglobin in rats during shock syndrome.

T2-selective proton imaging at 10 MHz is based on a Carr-Purcell-Meiboom-Gill pulse sequence with at least 36 echoes and 2 tau greater than or equal to 6 ms employing the projection reconstruction technique. The transverse magnetization decay of the volume elements is multiexponential composed of up to three monoexponential T2 components. The T2 values are arranged in T2 histograms which are slice specific. T2 windows are defined in the T2 histogram and used for imaging and for selecting T2 values which are assigned to proton classes and tissues. Tissue characterization is based on the visualization of differences in T2 behavior. The T2 values are transformed into 256 gray levels or, by use of a red-green-blue lookup table, into a rainbow code. Two display techniques have been developed and implemented in the RWTH Aachen Magnetic Resonance Software System (RAMSES): (i) for nonselective T2 imaging with secondary T2 selection and encoding of molecular classes on the basis of T2 values; (ii) for T2-selective imaging by employing combined primary and secondary T2 selection and encoding particular tissues and variations therein. The tissues can be identified by selecting different classes of molecules, like water, lipids, and proteins, when shifting the primary T2 window on the T2 axis of the T2 histogram.

Identification and characterization of tissues by T2-selective whole-body proton NMR imaging.

Two-dimensional T2-selective proton imaging of human head has been performed at 10 MHz employing Carr-Purcell-Gill-Meiboom pulse trains with echo separation of 6 and 12 msec. Using the information of 36 spin echoes and applying a recovery time of 2 sec the magnetization decay has been traced. The multi-exponential T2 relaxation curve of each pixel of the image has been deconvoluted into up to three mono-exponential functions which are defined by 1/T2,i and alpha i. The T2,i values are represented in a T2 histogram of the slice and then selected for generating images which relate to protons with specific T2. The alpha i values indicate the relative amounts of the T2-selected protons. Imaging on the basis of alpha i values increases the contrast of the image. The multiple T2-selective imaging technique leads to head images which show selectively cerebrospinal fluid, gray matter, different types of white matter, and nonassigned fast-relaxing proton classes.

Analysis of multiple T2 proton relaxation processes in human head and imaging on the basis of selective and assigned T2 values

Proton nuclear nagnetic resonance characterization of heme disorder in monomeric insect hemoglobins.

At room temperature aqueous solutions of dextrans with concentrations > 25% (w/w) exhibit a sol-gel transition in the presence of > 1.0 M potassium chloride. In dextrans the gelation was unexpected due to missing anionic groups that usually provide the binding sites for cations. The quantitative investigation of the gel formation is based on changes of the diffusibility of water and dextran chains. The apparent diffusion coefficients of bulk water (in the order of 10(-6) cm2/s) and of water trapped in the junction zones as well as of polymer chains (in the order of 10(-7) to 10(-8) cm2/s) are determined by employing pulsed field gradient stimulated echo (PFGSTE) NMR. The restricted diffusion of bulk water in viscous sols and in soft and rigid gels has been quantitatively analyzed providing data for interbarrier distances (pore size), permeabilities of the diffusion barriers (density of junction zones) and interbarrier diffusion coefficients of water. Based on already published x-ray structure data and in accordance with the diffusion data presented in this paper "potassium-bonding" is assumed to be the most important interaction for the formation of a microstructure and for the stabilization of cross-links. The ionic radius of the potassium ion perfectly fits to the cage established by six oxygen atoms of glucose units of three polymer chains. Other cations, such as Li+, Na+, Rb+ and Cs+, according to their nonfitting ionic radii, do not provoke dextran gelation under these conditions. The mechanism of the transitions from sol to soft gel and further to rigid gel is discussed on the basis of restricted diffusion and x-ray structure data.

Klaus Gersonde

Papers

Formation of free radicals and nitric oxide derivative of hemoglobin in rats during shock syndrome.

Identification and characterization of tissues by T2-selective whole-body proton NMR imaging.

Analysis of multiple T2 proton relaxation processes in human head and imaging on the basis of selective and assigned T2 values

Proton nuclear nagnetic resonance characterization of heme disorder in monomeric insect hemoglobins.

NMR studies on water and polymer diffusion in dextran gels. Influence of potassium ions on microstructure formation and gelation mechanism