Conformational stability of ribonuclease T1. I. Thermal denaturation and effects of salts.

The thermal transition of RNase T1 was studied by two different methods; tryptophan residue fluorescence and circular dichroism. The fluorescence measurements provide information about the environment of the indole group and CD measurements on the gross conformation of the polypeptide chain. Both measurements at pH 5 gave the same transition temperature of 56 degrees C and the same thermodynamic quantities, delta Htr (= 120 kcal/mol) and delta Str (= 360 eu/mol), for the transition from the native state to the thermally denatured state, indicating simultaneous melting of the whole molecule including the hydrophobic region where the tryptophan residue is buried. Stabilization by salts was observed in the pH range from 2 to 10, since the presence of 0.5 m NaCL caused an increase of about 5 degrees C to 10 degrees C in the transition temperature, depending on the pH. The fluorescence measurements on the RNase T1 complexed with 2'-GMP showed a transition with delta Htr =167 kcal/mol and delta Str =497 eu/mol at a transition temperature about 6 degrees C higher than that for the free enzyme. The large value of delta Htr for RNase T1 indicates the highly cooperative nature of the thermal transition; this value is much higher than those of other globular proteins. Analysis of the CD spectrum of thermally denatured RNase T1 suggests that the denatured state is not completely random but retains some ordered structures.     

蚕氨基酸代谢的研究——家蚕与蓖麻蚕的氨基酸和形成甘氨酸、丙氨酸的酶系比较及酮丙二酸的脱羧作用

家蚕(Bombyx mori L.)与蓖麻蚕(Samia,ricini B)的丝蛋白氨基酸含量差异甚为显著;家蚕丝含量最多的为甘氨酸(42.8％),其次为丙氨酸(32.4％);而蓖麻蚕丝则以两氨酸含量为最高(50.5％),其次为甘氨酸(27.8％)(Kirimuru等,1962)。为了阐明丝蛋白主要氨基酸的形成机制,我们曾报道了这两种蚕丝腺中,自L-天门冬氨酸与α-酮戊二酸形成丙氨酸的机制(许延森等,1964a);体液乙醛酸及丙酮酸与丝蛋白相应氨基酸的含量之     

Flexibility of bovine pancreatic trypsin inhibitor

The native conformation of a protein may be expressed in terms of the dihedral angles, phi's and psi's for the backbone, and kappa's for the side chains, for a given geometry (bond lengths and bond angles). We have developed a method to obtain the dihedral angles for a low-energy structure of a protein, starting with the X-ray structure; it is applied here to examine the degree of flexibility of bovine pancreatic trypsin inhibitor. Minimization of the total energy of the inhibitor (including nonbonded, electrostatic, torsional, hydrogen bonding, and disulfide loop energies) yields a conformation having a total energy of -221 kcal/mol and a root mean square deviation between all atoms of the computed and experimental structures of 0.63 A. The optimal conformation is not unique, however, there being at least two other conformations of low-energy (-222 and -220 kcal/mol), which resemble the experimental one (root mean square deviations of 0.66 and 0.64 A, respectively). These three conformations are located in different positions in phi, psi space, i.e., with a total deviation of 81 degrees, 100 degrees and 55 degrees from each other (with a root mean square deviation of several degrees per dihedral angle from each other). The nonbonded energies of the backbones, calculated along lines in phi, psi space connecting these three conformations, are all negative, without any intervening energy barriers (on an energy contour map in the phi, psi plane). Side chains were attached at several representative positions in this plane, and the total energy was minimized by varying the kappa's. The energies were of approximately the same magnitude as the previous ones, indicating that the conformation of low energy is flexible to some extent in a restricted region of phi, psi space. Interestingly, the difference delta phi i+1 in phi i+1 for the (i + 1)th residue from one conformation to another is approximately the same as -delta psi i for the ith residue; i.e., the plane of the peptide group between the ith and (i + 1)th residues re-orient without significant changes in the positions of the other atoms. The flexibility of the orientations of the planes of the peptide groups is probably coupled in a cooperative manner to the flexibility of the positions of the backbone and side-chain atoms.     

Non-polymerizable tropomyosin and control of the superprecipitation of actomyosin.

Non-polymerizable tropomyosin was prepared by the digestion of several C-terminal residues of tropomyosin with carboxypeptidase A [EC 3.4.12.2]. The intrinsic viscosity and molecular weight of the non-polymerizable tropomyosin were almost the same as those of untreated tropomyosin. Like untreated tropomyosin, the non-polymerizable tropomyosin in combination with troponin repressed the superprecipitation of actomyosin in the absence of calcium, while this repression was released by addition of calcium. However, the curve representing the superprecipitation rate as a function of pCa was less steep than that found with actomyosin containing untreated tropomyosin: in the former case, the rate increased to a plateau over about 2 pCa units, while in the latter case, it did so over about 1 pCa unit. These experimental results provide evidence that the "co-operation" in the regulation mechanism of skeletal muscle contraction, which is indicated by the steep curve of the contraction versus pCa relation, is mediated by tropomyosin-tropomyosin interaction along the thin filament.     

Allergenic glaucans from dermatophytes. II. Enzymic degradation

The action of pyruvic acid on glycerol leads principally to two isomeric, bicyclic lactones 1 and 2; this reaction is compared with that employing methyl and ethyl esters of pyruvic acid. The action of pyruvaldehyde or 2,3-butanedione on glycerol leads to bicyclic heterocycles having a secondary (5) or tertiary (6) hemiacetal group, respectively. These compounds, and likewise the derived alkoxy, acetoxy, and chloro analogs, are subject to the anomeric effect, the endo isomers being thermodynamically more stable than the exo isomers.     

Detection of somaclonal variations in tissue cultured date palm (Phoenix dactylifera L.) using transposable element-based markers

Plant Cell, Tissue and Organ Culture (PCTOC) - In vitro regeneration of date palm (Phoenix dactylifera L.) plants through somatic embryogenesis leads to the generation of somaclonal variants. The...