Expression of functional coagulation factor XIII in Escherichia coli.

Coagulation factor XIII is a zymogen that can be activated by thrombin cleavage to a transglutaminase that catalyses the formation of covalent crosslinks between fibrin chains in the final stages of the blood clotting cascade. Although circulating factor-XIII is composed of A and B subunits the catalytic activity is a property of the A subunits. In this study we have constructed a plasmid (pKKF13A) that contains a cDNA encoding the A subunit positioned downstream of a tac promoter.

Evolution of human alpha 1-acid glycoprotein genes and surrounding Alu repeats.

There is a mosaic pattern of variation between the two tandemly arranged human alpha 1-acid glycoprotein genes. Both the synonymous and the nonsynonymous sites of exons 3 and 4 are more divergent than the rest of the gene, suggesting that they have had a different evolutionary history. Comparisons of the two gene sequences with rat AGP indicate that exons 3 and 4 of AGP2 have been evolving without functional constraint since their divergence from AGP1.

Localization of the human UbB polyubiquitin gene to chromosome band 17p11.1-17p12.

The chromosomal location of the human ubiquitin genes has been evaluated by in situ hybridization. Because of the conservation of the ubiquitin sequence, coding-region probes cannot distinguish between specific ubiquitin genes and reveal ubiquitin sequences in a number of different chromosomal regions. The major sites of hybridization with a coding-region probe include 17p11.

Isolation and use of chromosome 1 probes for linkage studies on Charcot-Marie-Tooth disease.

Nine probes were isolated from a human chromosome 1 enriched library and mapped to regions of chromosome 1 using somatic cell hybrid lines. One clone, LR67, which mapped to 1q12----q23 detected a BglI RFLP. This probe, as well as 4 other known chromosome 1 markers, alpha-spectrin, Factor XIIIB, DR10 and DR78, were used for linkage studies in 15 Charcot-Marie-Tooth disease (CMT1) families.

Molecular genetics of the human glutathione S-transferase.

Multiple human cytosolic glutathione transferases have been described. These enzymes are the products of multiple genes that can be classified into at least four evolutionary classes. The genes encoding each class appear to be clustered on distinct chromosomes.

Genetic heterogeneity of the human glutathione transferases: a complex of gene families.

The glutathione transferases (GSTs) are involved in the metabolism of a wide range of compounds of both exogenous and endogenous origin. There is evidence that deficiency of GST may increase sensitivity to certain environmentally derived carcinogens. In contrast, elevated expression has been implicated in resistance to therapeutic drugs.

HLA A1, B8, DR3 extended haplotypes in autoimmune chronic hepatitis.

Genetic determinants of the autoimmune type of chronic active hepatitis include the major histocompatibility complex alleles HLA-B8 and HLA-DR3, which are usually present as the haplotype A1, B8, DR3. In certain other autoimmune diseases, an extended haplotype including complement alleles confers a greater relative risk than does B8, DR3. Hence, extended haplotypes were ascertained in autoimmune chronic active hepatitis by typing for HLA, complement alleles C4A, C4B, and Bf, and glyoxalase type 1 or 2.

Crystallization of GST2, a human class alpha glutathione transferase.

Single crystals of human GST2, a class alpha glutathione transferase have been grown in polyethylene glycol 2000 by the hanging-drop vapour diffusion method. The crystals belong to space group C2 and have cell dimensions a = 100.8 A, b = 95.

Isolation of a cDNA clone and localization of the human glutathione S-transferase 3 genes to chromosome bands 11q13 and 12q13-14.

A partial cDNA clone of the glutathione S-transferase 3 gene (GST3) was obtained by screening a lambda gt11 human lung cDNA library with antiserum to human lung GST3. The sequence of this cDNA showed two base differences from the coding sequence of a GST3 cDNA isolated from a human placental cDNA library. Hybridization of the cloned GST3 cDNA to human chromosomes resulted in a primary peak of grains over band 11q13, a localization predicted by prior experiments.

Unequal crossover generates variation in ubiquitin coding unit number at the human UbC polyubiquitin locus.

An observed mRNA length polymorphism of the human UbC polyubiquitin gene transcript was shown to correlate exactly with a three-allele HaeIII RFLP. Both polymorphisms apparently result from a variation in the number of ubiquitin coding units per UbC allele. Transcriptionally active alleles containing seven, eight, or nine coding units were observed, at frequencies suggesting that alleles of higher coding-unit number are selectively retained in the population.

The B subunit of coagulation factor XIII is linked to renin and the Duffy blood group to alpha-spectrin on human chromosome 1.

Family linkage studies were used to detect two linkage relationships on human chromosome 1. The B subunit of coagulation factor XIII showed significant linkage to renin with a maximum lod score of 5.071 at a distance of 10 cM.

Localization of the coagulation factor XIII B subunit gene (F13B) to chromosome bands 1q31-32.1 and restriction fragment length polymorphism at the locus.

In situ hybridization of tritiated cDNA probes for the gene for the B subunit of coagulation factor XIII localized the F13B locus to bands q31-q32.1 on human chromosome 1 and perhaps more precisely to sub-bands 1q31.2 or 1q31.

Molecular and serological characteristics of the glutathione S-transferases of Schistosoma japonicum and Schistosoma mansoni.

When aqueous extracts of Schistosoma japonicum and S. mansoni adult worms are passed over columns of glutathione-conjugated agarose, two molecular species of Mr 26,000 and Mr 28,000 are detected in eluates as analysed by SDS-PAGE, these eluates having glutathione S-transferase (GST) activity. The molecules, termed Sj26 and Sj28 from S.

Glutathione S-transferases in Fasciola hepatica.

Glutathione S-transferases (GST's) are widespread in the tissues of the liver fluke, Fasciola hepatica, and consist of multiple isozymes. Following purification to apparent homogeneity by affinity chromatography on glutathione agarose, fluke GST's were shown to comprise 2 components with molecular weights of about 25,000. Fluke GST's were immunogenic to rats, but when used as a vaccine conferred no protection on the animals against a challenge infection with F.

Structure and characterisation of a duplicated human alpha 1 acid glycoprotein gene.

Human alpha 1-acid glycoprotein (AGP), also known as orosomucoid, is a major acute-phase plasma protein. The amino acid sequence of AGP, which was determined by sequencing from protein isolated from pooled plasma, contained amino acid substitutions in 21 different positions. Genomic and cDNA clones which correspond to one of the possible amino acid sequences have been previously reported.

Haplotypes of the coagulation factor XIII A subunit locus in normal and deficient subjects.

Several RFLPs have been detected using a cDNA fragment encoding the amino-terminal half of the A subunit of factor XIII. The RFLPs show little linkage disequilibrium and form many different haplotypes that can be used to identify chromosomes transmitting factor XIII A subunit deficiency. Southern blot analysis of three deficient individuals from two families showed that, in these cases, factor XIII A subunit deficiency did not result from a major gene deletion or rearrangement.

Human muscle glutathione S-transferase (GST-4) shows close homology to human liver GST-1.

Human muscle specific glutathione S-transferase (RX: glutathione R-transferase, EC 2.5.1.

Expression of an enzymatically active parasite molecule in Escherichia coli: Schistosoma japonicum glutathione S-transferase.

The NH2-terminal amino acid sequence of the Mr 26 000 glutathione S-transferase (EC 2.5.1.

Localization of human alpha 1 acid glycoprotein genes to 9q31----34.1.

There is evidence for more than one alpha 1 acid glycoprotein (alpha 1 AGP) gene, and they all appear to be in close proximity. In situ hybridization of the cloned human cDNA p alpha 1AGP-2 to human chromosomes indicates that the alpha 1AGP genes are located between bands q31 and q34.1 on chromosome 9.

Localization of the coagulation factor XIII A subunit gene (F13A) to chromosome bands 6p24----p25.

Electrophoretic studies of the genetic variation of the A subunit of coagulation factor XIII (F13A) have shown that the A subunits expressed in placenta and in the circulation are the products of the same gene locus. In situ hybridization studies with a cDNA fragment encoding the amino terminal region of the A subunit have localized the gene to bands p24----p25 on chromosome 6. This confirms previous studies that showed linkage between F13A and the major histocompatibility complex.

Expression of human glutathione S-transferase 2 in Escherichia coli. Immunological comparison with the basic glutathione S-transferases isoenzymes from human liver.

A plasmid, termed pTacGST2, which contains the complete coding sequence of a GST2 (glutathione S-transferase 2) subunit and permits the expression of the protein in Escherichia coli was constructed. The expressed protein had the same subunit Mr as the enzyme from normal human liver and retained its catalytic function with both GST and glutathione peroxidase activity. Antiserum raised against the bacterially synthesized protein cross-reacted with all the basic GST isoenzymes in human liver.