共查询到20条相似文献,搜索用时 15 毫秒
1.
The effect of a decrease inpH on the structural integrity of carmin has been monitored by a variety of biophysical techniques. The protein undergoes initial dissociation up topH 3.5–4.0 without any significant denaturation. Below thispH the process of dissociation and denaturation appears to be simultaneous. Further, in thepH range of 2.5–1.6 the protein reassociates to probably a different polymer resulting from possibly, an entropically driven hydrophobic interaction. The process of dissociation appears to be reversible to a large extent. The process of denaturation appears to be governed by the kinetic path that the denatured protein molecule follows either by a sudden decrease inpH or through a gradual decrease inpH. These results are interpreted while keeping in view the oligomeric and globular structure of carmin at neutralpH. The results would help in understanding of structure-function relationship of the protein and its role in hydrogen ion bindingin vivo. 相似文献
2.
The effect of alkalinepH on the association, dissociation, and denaturation of carmin, the high-molecular-weight protein from safflower seed was investigated in thepH range 7–12, using various biophysical techniques. The results indicate that the multimeric protein carmin dissociates atpH 8.0 where denaturation has not set in. The association-dissociation of the protein can be represented schematically as 11S 7S 4S 2S. AbovepH 10, the protein undergoes simultaneous dissociation and denaturation. The denaturation process appears to be complete at pH 12.5. The protein undergoes conformational change and covalent modifications and cleavage during the denaturation process. A reversibility study shows that the process of dissociation is reversible to a large extent, whereas denaturation appears to be irreversible. These results are discussed in terms of association-dissociation, denaturation and alkaline-catalyzed covalent modifications and cleavage of seed proteins. 相似文献
3.
John Gutknecht 《The Journal of membrane biology》1984,82(1):105-112
Summary A simple method of measuring proton/hydroxide conductance (G
H/OH) through planar lipid bilayer membranes is described. First the total conductance (G
m
) is measured electrically. Then the H+/OH– transference number (T
H/OH) is estimated from the diffusion potential (V
m
) produced by a transmembrane pH gradient. The pH gradient is produced by a pair of buffered solutions which have identical concentrations of all ions except H+ and OH–. Thus,V
m
is due entirely to H+/OH– diffusion andG
H/OH can be calculated from the relations,V
m
=T
H/OH
E
H/OH andG
H/OH=T
H/OH
G
m
, whereE
H/OH is the equilibrium potential for H+ and OH–. In bilayers made from bacterial phosphatidylethanolamine (PE) inn-decane,G
H/OH is nearly independent of pH, ranging from about 10–9 S cm–2 at pH 1.6 to 10–8 S cm–2 at pH 10.5. BecauseG
H/OH is nearly independent of pH, the calculated permeability coefficients to H+ and/or OH– are extremely pH dependent, which partly explains the wide range of values reported for phospholipid vesicles and biological membranes.G
H/OH appears to be independent of the membrane surface charge, because titrating either the phosphate or the amino group of PE has little effect onG
H/OH.G
H/OH is reduced about 10-fold when the water activity is reduced 33% by replacement with glycerol. Although the mechanism of H+/OH– conductance is not known, the relation betweenG
H/OH and water activity suggests that several water molecules are involved in the H+/OH– transport process. 相似文献
4.
Bradycardial response inAplysia exposed to air 总被引:1,自引:1,他引:0
R. Feinstein H. Pinsker M. Schmale B. A. Gooden 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1977,122(3):311-324
Summary Heart rate was chronically monitored (Figs. 1, 3) in two species of the marine gastropodAplysia. The warm waterA. brasiliana have an average basal heart rate in water of 33 min–1, whereas the cold waterA. californica's heart rate is 20.6 min–1. The heart rate in both species shows a strong temperature dependence and the difference in basal heart rate is negligible when measured at the same temperature (Fig. 2). Both species show a consistent bradycardia when exposed to air (Fig. 4):A. brasiliana showed a 43% average decrease in air, whereasA. californica showed only a 16.5% decrease. Removal of the abdominal ganglion produced no significant decrease in heart rate in either species, nor did it reduce the bradycardial response to air exposure inA. californica (Fig. 8). However, it significantly reduced, but did not abolish, the bradycardia inA. brasiliana (Figs. 5, 6, 7). We conclude that the bradycardia has a significant central component inA. brasiliana, but is peripherally mediated inA. californica. The bradycardial response to air exposure may be analogous to the diving response in air breathing vertebrates. 相似文献
5.
Summary Weak to severe deficit of GATC sequences in the DNA of enterobacteriophages appears to be correlated with their undermethylation during growth indam
+ (GATC ade-methylase) bacteria. This observation is corroborated by the sequence analysis showing no evidence for site-specific mutagenicity of 6meAde. The MutH protein of the methyl-directed mismatch repair system recognizes and cleaves the undermethylated GATC sequences in the course of mismatch repair. To enquire whether the MutH function of the methyldirected mismatch repair system participates in counterselection of GATC sequences in enterobacteriophages, we have studied the yield of bacteriophage X174 containing either 0, 1, or 2 GATC sequences, in wild type,dam, andmut (H, L, S, U) Escherichia coli. Following transfection with unmethylated DNA containing two GATC sequences, a net decrease in the yield of infective particles was observed in all bacterialmutH
+ dam– strains, whereas no detectable decrease was observed in bacteria infected by DNA without GATC sequence. This effect of the MutH function is maximum in wild type andmutL andmutS bacteria whereas the effect is not significant inmutU bacteria, suggesting an interaction of the, helicase II with the MutH protein.However, indam
+ bacteria, the presence of GATC sequences leads to an increased yield of infective particles. The effect of GATC sequence and its Dam methylation system on phage yield inmutH
– bacteria reveals that methylated GATC sequences are advantageous to the phage. These results suggest that the methyl-directed mismatch repair system, and in particular its MutH protein, may have participated in severe counterselection of GATC sequences from enterobacteriophages, presumably, by DNA cleavage or by interfering with DNA replication or packaging when GATC sequences are undermethylated. Coevolution of the Dam and MutH proteins could then account for the loss of GATC sequences from DNA of bacteriophages growing indam
+ hosts. 相似文献
6.
Jun-ling You Saskia C. F. Milton Raymond C. deLisle Milton N. S. Rangaraju Robert B. Harris 《Journal of Protein Chemistry》1993,12(2):133-141
Homogeneous pre-pro-GnRH/GAP protein was recently synthesized in 100 mg quantities by solid-phase methods and surprisingly, the synthetic pre-pro-protein, which normally does not escape the endoplasmic recticulum, was found to inhibit the release of prolactin from cultured pituitary cells. This is the first demonstration of significant biological activity associated with a precursor protein and provides the rationale for its further study. We now report the results of our initial examination of the conformational properties of pre-pro-GnRH/GAP protein as a prelude to solving its solution phase conformation by homonuclear1H-NMR protocols. Thermal andpH titration fluorescence and circular dichroism spectroscopies reveal that the protein is resistant to thermal-induced conformational changes but is particularly sensitive topH-induced conformational changes; while Asp/Glu and Arg residues may contribute to structural stability, His and Lys residues predominate. Pre-pro-GnRH/GAP is about 30% helix in the range of 2–40°C; however, even at 90°C, the peptide retains nearly 50% of its helix character. There is no evidence for a cooperative transition; for this reason, differential scanning calorimetry failed to yield a defined transition thermogram. Pre-pro-GnRH/GAP apparently does not pass through a transition state as a function of temperature but appears to flex and retain a high percentage of helix structure, resulting in subtle changes in secondary structure. There is no discernible isodichroic point. On either side of the neutralpH range, however, there are dramatic changes in structure that result in nonreversible denaturation of the protein. Relative to N(Ac)Trp-amide, the emission position of intrinsic Trp fluorescence of pre-pro-GnRH/GAP is blue shifted to 338 nm, indicating that the microenvironment(s) encompassing the 2 Trp residues are buried within the protein structure. Synthetic pre-pro-GNRH/GAP is a substrate for GAP-releasing enzyme (the proposed physiologically relevant processing enzyme of the precursor protein) and yields GAP peptide (D14–I69). Of the other serine proteinases tested (trypsin, plasmin, kallikrein), only GAP-releasing enzyme shows this specificity of cleavage. Hierarchical cleavage observed in the time course of proteolysis with trypsin, however, suggests that other peptide products might be formed from GAP once it is processed from the precursor protein by cleavage at sites other than the primary processing site catalyzed by enzymes other than GAP-releasing enzyme. The primary processing site for GAP-releasing enzyme (GLRPGGKR) is thus accessible in the precursor protein, consistent with our hypothesis that the recognition sequence is located at the surface of the protein and acts as a recognition element for the processing endoproteinase. The conformation of the precursor protein is dynamic, supporting the idea that intracellular (and/or intragranular) conditions may play a role in regulation of endoproteolysis. Conformational flexing of the pro-hormone in response to intracellular conditions may serve to differentially expose various processing sites which may help explain tissue specificity of processing. 相似文献
7.
Purification,electrophoretic behavior,and kinetics of iron release of liver ferritin of Dasyatis akajei 总被引:5,自引:0,他引:5
Kong B Huang HQ Lin QM Kim WS Cai Z Cao TM Miao H Luo DM 《Journal of Protein Chemistry》2003,22(1):61-70
From the liver of fish Dasyatis akajei, ferritin has been isolated by thermal denaturation and ammonium sulfate fractionation and then further purified by anion exchange chromatography and gel exclusion chromatography. The molecular weight of the liver ferritin of D. akajei (DALF) was measured to be 400 kDa by PAGE. Moreover, SDS-PAGE experimentation indicates that protein shell of DALF consists of the H and L subunits with molecular weight of 18 and 13 kDa, respectively. Using isoelectric focusing with pH ranging from 5.0 to 6.0, the ferritin purified by the PAGE exhibited three bands with different pI values in the gel slab. Diameters of the protein shell and iron core were also investigated by transmission electron microscope and determined to be 10–12 nm and 5–8 nm, respectively. A kinetic study of DALF reveals that the rate of self-regulation of the protein shell rather than the complex surface of the iron core plays an important role in forming a process for iron release with mixed orders. 相似文献
8.
Tadeusz Pawelczyk Renata Kowara Filip Golebiowski Andrzej Matecki 《Protein expression and purification》2000,18(3):320
The fragile histidine triad (Fhit) protein is a homodimeric protein with diadenosine 5′,5-P1,P3-triphosphate (Ap3A) asymmetrical hydrolase activity. We have cloned the human cDNA Fhit in the pPROEX-1 vector and expressed with high yield in Escherichia coli with the sequence Met-Gly-His6-Asp-Tyr-Asp-Ile-Pro-Thr-Thr followed by a rTEV protease cleavage site, denoted as “H6TV,” fused to the N-terminus of Fhit. Expression of H6TV–Fhit in BL21(DE3) cells for 3 h at 37°C produced 30 mg of H6TV–Fhit from 1 L of cell culture (4 g of cells). The H6TV–Fhit protein was purified to homogeneity in a single step, with a yield of 80%, using nickel-nitrilotriacetate resin and imidazole buffer as eluting agent. Incubation of H6TV–Fhit with rTEV protease at 4°C for 24 h resulted in complete cleavage of the H6TV peptide. There were no unspecific cleavage products. The purified Fhit protein could be stored for 3 weeks at 4°C without loss of activity. The pure protein was stable at −20°C for at least 18 months when stored in buffer containing 25% glycerol. Purified Fhit was highly active, with a Km value for Ap3A of 0.9 μM and a kcat(monomer) value of 7.2 ± 1.6 s−1 (n = 5). The catalytic properties of unconjugated Fhit protein and the H6TV–Fhit fusion protein were essentially identical. This indicates that the 24-amino-acid peptide containing the six histidines fused to the N-terminus of Fhit does not interfere in forming the active homodimers or in the binding of Ap3A. 相似文献
9.
Summary Basal lateral membrane vesicles were isolated from rat intestinal epithelial cells. The sodium potassium triphosphatase (Na/K-ATPase) of these plasma membranes has been characterized by (1) the molecular weight of the phosphorylated intermediate, (2) the sensitivity of the phosphorylated intermediate to hydroxylamine, (3) its ouabain binding constants, and (4) its susceptibility to digestion by pronase. The phosphorylated intermediate was shown by SDS polyacrylamide gel electrophoresis to be a protein of 100,000 Daltons apparent mol wt. Its extensive hydrolysis in hydroxylamine demonstrated that it was an acyl phosphate. The isolated basal lateral membranes bound ouabain with a dissociation constant,K
m
(1.5×10–5
m), similar to the inhibitory constantK
I
(3×10–5
m), measured for ouabain inhibition of the Na/K-ATPase activity. The association rate constant measured for ouabain binding at 22°C was 1.3×103
m
–1 sec–1 and is similar to the association rate constants reported for other tissues and species. The high dissociation rate constant, 3.6×10–2 sec–1, is consistent with the insensitivity of the rat to ouabain. Digestion of the intact cells by pronase yielded basal lateral membranes in which the Na/K-ATPase had been unaffected. The phosphorylated intermediate ran as a sharp band at 100,000 Daltons on electrophoresis, and the ouabain dissociation constant appeared to be unchanged. In these membranes, protein stains of polyacrylamide gels revealed digestion of the major high mol wt proteins including the major protein at 100,000 Daltons. This suggests that the Na/K-ATPase represent a minor component, less than 1%, of the basal lateral membrane protein. From these characteristics of the phosphorylated intermediate and the ouabain binding constants, we conclude that the Na/K-ATPase of the basal lateral membranes of rat intestinal epithelial cells is similar to that found in other tissues and species. Estimates of the number of pump sites and the turnover number predict rates of Na transport that are consistent with observed values.This paper is dedicated to the memory of Professor David H. Smyth, FRS, who died on September 10, 1979. 相似文献
10.
A low-Mr tight binding proteinase inhibitor was purified from bovine muscle by alkaline denaturation of cysteine proteinases, gel filtration on Sexphadex G-75 and affinity chromatography on carboxymethyl-papain-Sepharose. Chromatofocusing separated three isoforms which are similar in their Mr of about 14 000, their stability with heating at 80°C and their inhibitory activity towards cathepsin H, cathepsin B and papain. The equilibrium constants (Ki) were determined for these three cysteine proteinases but for cathepsin H, association (kass) and dissociation (kdiss) rate constants were also evaluated. Ki values of 56 nM and 8.4 nM were found for cathepsin B and cathepsin H, respectively. For papain, Ki was in the range of 0.1–1 nM. The kinetic features of enzyme-inhibitor binding suggest a possible role for this low-Mr protein inhibitor in controlling ‘in vivo’ cathepsin H proteolytic activity. With regard to cathepsin B, such a physiological role was less evident. 相似文献
11.
Suara A. Adediran 《Journal of Protein Chemistry》1984,3(5-6):437-444
The formation of Compounds II and III of horseradish peroxidase from Compound I and potassium ferrocyanide and from Compound II and excess hydrogen peroxide, respectively, was studied as a function ofpH at 25°C and a constant ionic strength of 0.11. The yield of Compound II obtained increases progressively with increase inpH; a mixture of Compounds I and II is produced at acidicpH. Pure Compound III is obtained at allpH values, but the highest yield is obtained atpH values between 6.0 and 7.0. The yield of p-670, formed when Compound III is allowed to stand for 60 min, decreases with increase inpH, while the decay of Compound III also decreases with increase inpH. Therefore p-670 is the decay product of Compound III. 相似文献
12.
The structural stability of phaseolin was determined by using absorbance, circular dichroism (CD), fluorescence emission, and fluorescence polarization anisotropy to monitor denaturation induced by urea, guanidinium chloride (GdmCl),pH changes, increasing temperature, or a combination thereof. Initial results indicated that phaseolin remained folded to a similar extent in the presence or absence of 6.0 M urea or GdmCl at room temperature. In 6.0 M GdmCl, phaseolin denatures at approximately 65°C when probed with absorbance, CD, and fluorescence polarization anisotropy. The transition occurs at lower temperatures by decreasingpH. Kinetic measurements of denaturation using CD indicated that the denaturation is slow below 55°C and is associated with an activation energy of 52 kcal/mol in 6.0 M GdmCl. In addition, kinetic measurement using fluorescence emission indicated that the single tryptophan residue was sensitive to at least two steps of the denaturation process. The fluorescence emission appeared to reflect some other structural perturbation than protein denaturation, as fluorescence inflection occurred approximately 5°C prior to the changes observed in absorbance, CD, and fluorescence polarization anisotropy. 相似文献
13.
M. Bornancin G. De Renzis J. Maetz 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1977,117(3):313-322
Summary Freshwater eel gills are notorious for their limited ability to pump chloride. As a result there is a considerable discrepancy between the Na+ and Cl– plasma levels, and plasma HCO3
– and blood pH are relatively high in this species.When eels are kept in tanks aerated with pure oxygen, significant alterations in blood acid-base balance, an increase in plasma pCO2 and a decrease in blood pH, are observed. In fish studied after 3 weeks hyperoxia, the decrease in blood pH is compensated by an increase in plasma HCO3
–. Such fish exhibit a Cl– influx 5 times higher than that observed in normoxic fish. This Cl– influx is readily inhibited by addition of SCN– to the external medium.An anion-stimulated ATPase activated by HCO3
– and by Cl– and inhibited by SCN– was recently described in membrane fractions of the gills ofCarassius auratus, a fish noted for its high Cl– pumping rate. This enzyme is also found in the gills of the eel. While the maximal rates of enzyme activation by HCO3
– and by Cl– are similar inCarassius andAnguilla, the affinity of the enzyme for Cl– is 25 times higher inCarassius. In the microsomal fraction of the hyperoxic eel gills, the maximal anionstimulated ATPase activity remains unchanged but HCO3
– affinity decreases by 50%, while Cl– affinity increases 5 times. Thus some characteristics of this ATPase seem to be closely related to the Cl– pump activity exhibited by the gill in fresh water. 相似文献
14.
Estimates of nuclear DNA base composition by determination of thermal denaturation temperatures (Tm) indicate guanine + cytosine (G + C) levels of 35.4–46.8% for ten species of the Gracilariaceae, representing the generaGracilaria andHydropuntia. Tm values were found to be reproducible with variation among most samples and replicates of less than 1 °C and 2 mol%. Interspecific variation in G + C values was less than 11.4% amongGracilaria species. Calculation of intragenomic base pair composition distribution based on mid-resolution thermal denaturation (A 1 °C/min with 4s interval H and dT logging) indicated an inverse relationship between maximum similarity values and taxonomic rank. Intraspecific (population level) maximum similarity (homology) values were estimated to range from 79–90% inGracilaria tikvahiae (4 isolates). Interspecific values of 46–69% were found in 13 species ofGracilaria. Nucleotide distribution similarity values for the Gracilariaceae are compared with previous information for genome organization and complexity, genome size and karyotype patterns.Author for correspondence 相似文献
15.
Mononucleosomes containing 143 ± 6 base pairs of duplex DNA and approximately two copies each of the histones H2a, H2b, H3 and H4 were examined during thermal denaturation by high resolution electron microscopy using both bright- and dark-field (tilted beam) modes. Co-operative destabilization and unfolding of the 13.2 ± 1.4 nm diameter toroids occurred only after the second of the two major melting transitions. The unfolding patterns are consistent with about 1.5–1.8 turns of supercoiled DNA in intact nucleosomes. The dominant unfolded structure of samples post-fixed with glutaraldehyde is a 17.5 ± 2.1 nm diameter open ring. Both sister DNA strands remain associated with protein. The distribution and shape of the protein patches are more irregular in unfixed, unstained samples visualized by darkfield microscopy. Image reconstruction studies on fixed and stained ring-shaped specimens indicates that there are 6–10 globular protein elements or patches, each about 3.9 ± 0.5 nm in diameter, per DNA moiety. 相似文献
16.
Inactivation of glucose 6-phosphate dehydrogenase (G6PDH) complexed with its substrate, glucose 6-phosphate (GP), or cofactor, NADP+, has been studied within the range 20–40°C in three media: (a) 0.04 M NaOH–glycine buffer (pH 9.1); (b) Aerosol OT (AOT) reversed micelles in octane; and (c) Triton X-100 micelles in octane supplemented with 10% hexanol. The enzyme inactivation was characterized quantitatively by first order rate constants, k
in(s–1). In the case of G6PDH–NADP+complexes, the values of k
inwere independent of the initial concentrations of G6PDH, either in aqueous medium or AOT micelles. The values of k
infor the complex G6PDH–GP were inversely related to the initial concentration of the enzyme, in both aqueous and micellar media. When inactivation of both complexes were studied in AOT micelles, minimum values of k
incorresponded to the degree of hydration W
0= 16.7; at W
0> 16.7 and W
0< 16.7, k
inincreased. Within the range 20–40°C, the values of k
inmeasured for both complexes in aqueous medium were significantly lower than those measured in AOT micelles. Temperature dependences of k
inwere characterized by inflections in Arrhenius plots, which corresponded, depending on the medium, to certain temperatures from 33.6°C to 40°C. In all media studied, NADP+complexes of the enzyme exhibited higher stability than their GP counterparts. The parameters of G6PDH and G6PDH–NADP+melting, measured by differential scanning microcalorimetry (maximum temperature and half-width of the transition, enthalpy of denaturation, and van't Hoff enthalpy), provided unequivocal evidence of the higher stability of the complex as compared to that of the enzyme. In addition, this approach demonstrated that G6PDH undergoes destabilization in AOT micelles. 相似文献
17.
Juli O'Connor Dean F. Revell Karen E. Masters Ian F. Connerton Ian G. Sumner 《Protein expression and purification》1996,7(4):377-383
The gene encoding the 67-kDa cocoa storage protein precursor has been cloned fromTheobroma cacaoand expressed inEscherichia coliusing the pET expression system. The recombinant storage protein has been renatured from inclusion bodies at 30°C using 20 m
glycine–NaOH buffer, pH 10.0, containing 1 m
oxidized glutathione and 0.1% Brij. The renatured protein was purified and demonstrated to adopt a stable native conformation by optical spectroscopy. Secondary structure analysis from circular dichroism indicated the protein to be 23% α-helix and 38% β-sheet, in close agreement with values obtained using a secondary structure prediction program. 相似文献
18.
Hoch FL 《Journal of bioenergetics and biomembranes》1976,8(5):223-238
Measurements of fluorescence at >420 nm and extracted NADPH in mitochondria obtained from the livers of hypothyroid rats show that the addition of Pi, ADP and glutamate rapidly reduces over 90% of the total reducible intrinsic pyridine nucleotides in State 3, compared with 20% in normals. The total fluorescence intensity change and reducible NADP+ is about twice normal in hypothyroid mitochondria. Adding 6–30 µMl-thyroxine to hypothyroid mitochondriain vitro decreases and delays the substrate-induced reduction of pyridine nucleotides, and excludes both NADP+ from such reduction and NADPH from oxidation by added ADP + Pi, without changing the high NADP(H) content. The correcting actions of the hormone are rapidly reversed by albumin, probably by binding free hormone. Changes in respiration do not appear to account for these observations. There is indirect evidence for decreased phosphorylation of added ADP in hypothyroid mitochondria, and a correction by added hormone. The hormonal actions on NADP(H) redox reactions are not reproduced by 1 to 6 µM dinitrophenolin vitro.l-Thyroxine appears to specifically block the participation of NADP (H) in redox reactions in mitochondria from hypothyroid rats, perhaps by effecting a sequestration of the nucleotide, by inhibiting the pyridine nucleotide transhydrogenase, or by activating an energy-linked process that competes with transhydrogenation.Papers I–III in this series were published inArch. Biochem. Biophys.I–124 (1968) 238.II–124 (1968) 248.III–150 (1972) 618.This work was supported by grants from the NIH (AM13564) and from The John A. Hartford Foundation. 相似文献
19.
The cyclic AMP receptor protein (CRP) regulates the expression of many genes in Escherichia coli. The protein is a homodimer, and each monomer is folded into two distinct structural domains. In this study, we have used differential scanning calorimetry (DSC) and circular dichroism (CD) to measure the enthalpy change and melting temperature of the apo-CRP and CRP complexes with cAMP or DNA sequences lac, gal, and palindromic ICAP. DSC and CD measurements showed irreversible thermal denaturation process of CRP. Enthalpy of dissociation of the protein–DNA complex, as measured by DSC, depends on the DNA sequence. The thermal transition of the protein in CRP-DNA complexes, measured by CD, indicates that the protein stability in the complex is also DNA sequence-dependent. 相似文献
20.
Bruno Jirgensons 《Journal of Protein Chemistry》1982,1(1):71-84
The factors determining the onset and extent of reconstructive denaturation of proteins were considered by comparing circular dichroism (CD) data of seven proteins and previously published findings. The effects of sodium dodecyl sulfate (SDS) on the conformation of the following proteins were tested: lysozyme, the mitogens fromPhytolacca americana (fractions Pa2 and Pa4), lectin fromWistaria floribunda, ovine lutropin, a Bence Jones protein, and histone H2B. While the helix content of lysozyme was raised by SDS slightly, in the Bence Jones protein andW. floribunda lectin it increased from near zero to about 25–30%. In histone H2B the helix content was raised by SDS even to about 48%. However, no clear indication of helix formation could be observed in the mitogens and lutropin, even at low pH or 2.0–2.5. The tertiary structure of the proteins was perturbed by SDS. It was concluded that the reorganization of secondary structure of the proteins was favored by the following factors: (1) presence of helicogenic amino acid sequences in the protein, (2) availability of positively charged sites of the basic amino acids for interactions with the dodecyl ion, (3) absence of a large surplus of negatively charged sites on the surface of protein, and (4) absence of extensive disulfide cross-linking within the macromolecule. Both hydrophobic and electrostatic interactions occur in reconstructive denaturation, and the newly formed helices are stabilized by hydrophobic shielding by the alkyl chains of the alkyl sulfate. 相似文献