Hydrophilic proteins

There is Httle difference between the wet and the dry stress—strain diagrams of hydrophobic fibers, eg, nylon, acryHc, and polyester. Hydrophilic protein fibers and regenerated cellulose exhibit lower tensile moduH on wetting out, that is, the elongations increase and the strengths diminish. Hydrophilic natural ceUulosic fibers, ie, cotton, linen, and ramie, are stronger when wet than when dry. 

Aqueous-detergent solutions of appropriate concentration and temperature can phase separate to form two phases, one rich in detergents, possibly in the form of micelles, and the other depleted of the detergent (Piyde and Phillips, op. cit.). Proteins distribute between the two phases, hydrophobic (e.g., membrane) proteins reporting to the detergent-rich phase and hydrophilic proteins to the detergent-free phase. Indications are that the size-exclusion properties of these systems can also be exploited for viral separations. These systems would be handled in the same way as the aqueous two-phase systems. 

Deactivation generally refers to a change in the physical structure of the enzyme, often caused by an increase in temperature. Some of the amino acids in a protein chain are hydrophobic. Others are hydrophilic. Proteins in solution fold into elaborate but characteristic shapes to increase like-to-like interactions... 

PelZ is a hydrophilic protein of 420 amino acids with a short hydrophobic sequence at its N-terminal end which has Ae characteristics of the signal sequences of exported proteins. The signal peptide may be 24 amino acids long, which would corroborate wiA the usual length encountered in prokaryotes. The molecular cloning of the pelZ gene in an expression vector pT7-6 allowed for the specific 35S-cysteine-methionine raAo-labelling of PelZ in E. coli K38. We could detect, in crude extracts, the presence of a precursor and a mature form of PelZ. After cell fractionation, Ae mature form of PelZ could be localized in Ae periplasm of E. coli. So PelZ appears to be a protein exported by Ae Sec-dependent system of translocation. 

Cervera and Levine [81] studied the mechanism of oxidative modification of glutamine synthetase from Escherichia coli. It was found that active oxygen species initially caused inactivation of the enzyme and generated a more hydrophilic protein, which still was not a substrate for the protease. Continuous action of oxygen species resulted in the formation of oxidized protein subjected to the proteolytic attack of protease. 

New insights into the analysis of hydrophobically post-translational modified proteins could be achieved by the construction of lipidated proteins in a combination of bioorganic synthesis of activated lipopeptides and bacterial expression of the protein backbone (Fig. 19). The physico-chemical properties of such artificial lipoproteins differ substantially from those of the corresponding lipopeptides. The pronounced dominance of the hydrophilic protein moiety (e.g., for the Ras protein 181 amino acids) over a short lipopeptide with one or two hydrophobic modifications provides solubility up to 10 4 mol/1, while the biotinylated or fluorescence labeled lipopeptides exhibit low solubility in aqueous solutions and can be applied in the biophysical experiments only in vesicle integrated form or dissolved in organic solvent. 

In the development of the protein-fatty acid condensates it was possible to combine the renewable resources fatty acids (from vegetable oil) and protein, which can be obtained from both animal waste (leather) as well as from many plants, to construct a surfactant structure with a hydrophobic (fatty acid) and a hydrophilic (protein) part (Fig. 4.12). This was carried out by reacting protein hydrolysate with fatty acid chloride under Schotten-Baumann conditions using water as solvent. Products are obtained that have an excellent skin compatibility and, additionally, a good cleaning effect (particularly on the skin) and, in combination with other surfactants, lead to an increase in performance. For instance, even small additions of the acylated protein hydrolysate improve the skin compatibility. An... 

Figure 9.15 Enzymes in aqueous (light-coloured) and hydrophobic (shaded) phases. (A) A protein in the periplasm (PP) of a cell (OM = outer membrane, CM = cytoplasmic membrane) (B) membrane-bound protein in a lipid bilayer (C) hydrophilic protein in an inverted micelle (D) interaction between enzyme and substrates in aqueous micelles (E) graph of catalytic activity as a function of micelle concentration.

If the virus is treated with proteolytic enzymes the fuzzy layer formed by the viral spikes is removed (Osterrieth, 1965 Compans, 1971 Gahm-berg et al, 1972 Sefton and Gaffney, 1974 Utermann and Simons, 1974). Remnants of both El and E2 are left in the bilayer. These have a hydrophobic amino acid composition, and are soluble in lipid solvents such as chloroform-methanol. The amphiphilic nature of the spike protein is also evident from its capacity to bind Triton X-100 (0.6 g/g protein) which binds to the hydrophobic part to form a water-soluble protein-detergent complex (Simons et al., 1973a). The ability of amphiphilic proteins to bind Triton can be used to separate them from hydrophilic proteins using an extraction procedure recendy described... 

Cloud point extraction from biological and clinical samples. The most frequent use of CPE is for the separation and purification of biological analytes, principally proteins. In this way, the cloud point technique has been used as an effective tool to isolate and purify proteins when combined with chromatographic separations. Most of the applications deal with the separation of hydrophobic from hydrophilic proteins, with the hydrophobic proteins having more affinity for the surfactant-rich phase, and the hydrophilic proteins remaining in the dilute aqueous phase. The separation of biomaterials and clinical analytes by CPE has been described [105,106,113]. 

This chapter aims to summarize our efforts to investigate the effects of fluorinated amino acid substitutes on the interactions with natural protein environments. In addition to a rather specific example concerning the interactions of small peptides with a proteolytic enzyme, we present a simple polypeptide model that aids for a systematic investigation of the interaction pattern of amino acids that differ in side chain length as well as fluorine content within both a hydrophobic and hydrophilic protein environment. Amino acid side chain fluoiination highly affects polypeptide folding due to steric effects, polarization, and fluorous interactions. 

The a-helical coiled coil-based screening system already provided a wide variety of information about the interactions of fluorinated amino acids within hydrophobic and hydrophilic protein environments. Investigations on the thermal stability as well as the replicase activity have both emphasized the orthogonal properties of fluorinated aliphatic amino acid side chains. The term orthogonal in this context has been chosen by us to demonstrate that they are in fact hydrophobic... 

Frost-resistant (hardy) plants are less sensitive than others to damage by low temperatures that is caused by water loss and intracellular, ice-crystal formation. Production of such highly hydrophilic proteins as glycoproteins would constitute a potential mechanism, through the formation of hydrogen... 

Hydrophobic dehydration results from bonding of the protein s hydrophobic patches to the hydrophobic regions on the adsorbent. The enthalpic part of this interaction is small the entropy change is positive. Hydrophobic dehydration is relatively unimportant for hydrophilic surfaces and/or rigid hydrophilic proteins. 

Hydrophobic interaction chromatography relies on hydrophobic interactions between apolar amino acid residues in the proteins and a resin containing hydrophobic groups, such as /7-octyl or phenyl groups. After the protein mixture is applied to the column, an elution buffer with decreasing ionic strength is used. Hydrophilic proteins will elute first, whereas hydrophobic proteins elute last. 

A different class of hydrophilic proteins is encoded by the rab (ABA responsive) genes. These genes were first discovered as ABA-induced... 

A plot of the temperatures required for clouding versus surfactant concentration typically exhibits a minimum in the case of nonionic surfactants (or a maximum in the case of zwitterionics) in its coexistence curve, with the temperature and surfactant concentration at which the minimum (or maximum) occurs being referred to as the critical temperature and concentration, respectively. This type of behavior is also exhibited by other nonionic surfactants, that is, nonionic polymers, // - a I k y I s u I Any lalcoh o I s, hydroxymethyl or ethyl celluloses, dimethylalkylphosphine oxides, or, most commonly, alkyl (or aryl) polyoxyethylene ethers. Likewise, certain zwitterionic surfactant solutions can also exhibit critical behavior in which an upper rather than a lower consolute boundary is present. Previously, metal ions (in the form of metal chelate complexes) were extracted and enriched from aqueous media using such a cloud point extraction approach with nonionic surfactants. Extraction efficiencies in excess of 98% for such metal ion extraction techniques were achieved with enrichment factors in the range of 45-200. In addition to metal ion enrichments, this type of micellar cloud point extraction approach has been reported to be useful for the separation of hydrophobic from hydrophilic proteins, both originally present in an aqueous solution, and also for the preconcentration of the former type of proteins. 

Figure 8. Diagrammatic illustration of the two-phase model which describes the stratum corneum as a hydrophilic protein gel dispersing in a continuous lipid matrix.

Tables VI and VII ). On the other hand, capric acid and decylmethyl sulfoxide showed a dual effect on the hydrophilic protein gel and also on the lipophilic fatty matrix. In the case of capric acid, the overall enhancement in the permeation of progesterone was increased by 354%, in which the protein gel pathway and fatty matrix pathway contribute approximately equally (with enhancement factor of 15.3 vs. 13.0). In the case of decylmethyl sulfoxide, the overall enhancement was improved by 515% (40.2 vs. 7.8).

---