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181.
Flounder muscle (Pseudopleuronectes americanus) glyceraldehyde-3-phosphate dehydrogenase was characterized as to its stability towards various inactivating treatments in the presence and absence of the enzyme cofactor, NAD. Incubation of a partially purified enzyme preparation at urea concentrations greater than 2 M produced a very rapid inactivation. NAD greatly reduced the rate of inactivation at all the urea concentrations tested. Incubation of each of the three major muscle enzyme forms in 0.1 percent trypsin or chymotrypsin for forty-five minutes decreased the activity of each form by 65 percent and 55 percent, respectively. NAD (5mM) afforded complete protection to each enzyme form from proteolytic digestion by these two enzymes. Exposure of each form to 50 degrees or 20 mM ATP also led to gross inactivation which could be greatly reduced if the respective incubations were performed in the presence of 5mM NAD. NAD was also found to be required for the renaturation of the unfolded urea-denatured subunits to form the active tetramer. 相似文献
182.
The molecular organization of the beta-globin complex of the deer mouse, Peromyscus maniculatus 总被引:1,自引:0,他引:1
Padgett RW; Loeb DD; Snyder LR; Edgell MH; Hutchison CA d 《Molecular biology and evolution》1987,4(1):30-45
Recombinant DNA clones have been isolated that contain 80 kb of the
beta-globin complex from the deer mouse, Peromyscus maniculatus.
Comparisons of this complex with that from the laboratory mouse, Mus
domesticus (with an order 5'-Hbby, Hbb-bhO, Hbb-bhl, Hbb-bh2, Hbb-bh3,
Hbb-bl, Hbb-b2 3') highlight organizational trends in the beta-globin
complex since the two species diverged. Unlike other mammals studied thus
far, the deer mouse possesses three adult genes. Partial sequence analysis
indicates that each of the three adult genes is intact and hence may be
functional. Hybridization of one of the two Mus pseudogenes, Hbb-bh3, to
genomic blots from Peromyscus reveals that it has a homologous counterpart
in Peromyscus. Homologous genes to the two gamma-like Mus genes, Hbb-bhO
and Hbb-bhl, are also found in Peromyscus. The strong hybridization between
the Hbb-bhl genes and significant nucleotide similarity between the Hbb-bhO
genes suggest that both pairs are important for the ontogeny of these mice
although no known product has been identified for the Hbb-bhO genes. The
presence of Hbb-bhO and Hbb-bhl in Peromyscus suggests that the duplication
that created this related gene set occurred before the two lineages
diverged. A single gene for Hbb-y has been isolated from Peromyscus. The
adult region in Peromyscus has undergone significant divergence from the
same region in Mus, having three rather than two adult genes, the
acquisition of at least 15 kb of extra DNA relative to Mus, and possibly
the loss of the Hbb-bh2 pseudogene. The nonadult region of the complex, in
contrast, contains the same set of genes apparently distributed over the
same amount of DNA as in the Mus beta- globin complex. This observation
suggests that the embryonic region of the complex is more evolutionarily
stable than the adult region.
相似文献
183.
Alkis Constantinides Wolf R. Vieth Peter M. Fernandes 《Molecular and cellular biochemistry》1973,1(1):127-133
Summary The enzyme glucose oxidase (E.C. 1.1.3.4) was immobilized on collagen — a proteinaceous material found in biological systems as a structural material for a wide variety of cells and membranes. The novel technique of electrocodeposition, which utilizes the principles of electrophoresis, was used to deposit the enzyme-collagen complex on stainless steel helical supports. This technique has been developed in our laboratory. The mechanism of complex formation between collagen and enzyme involves multiple salt linkages, hydrogen bonds and van der Waals interactions.As a first step toward examining its feasible technical use, the kinetic behavior of the collagen-supported glucose oxidase was studied in a batch recycle type reactor and was compared with that for the soluble form. A novel reactor configuration consisting of multiple concentric electrocodeposited helical coils was used. The reactor was found to attain a stable level of activity which was maintained for several months under cyclic testing. The optimum levels of pH and temperature for the immobilized form of the enzyme were the same as those of the soluble enzyme, but the immobilized enzyme was more active than the soluble form at higher temperatures and pH. The values of the Michaelis-Menten parameters indicate that the overall reaction rate of the immobilized enzyme may be partially restricted by bulk and matrix diffusion. 相似文献
184.