Restriction site polymorphism in genes encoding type 2 but not type 1 gonococcal IgAl proteases

Hydrogenophaga intermedia strain S1 and Agrobacterium radiobacter strain S2 form a mixed bacterial culture which degrades sulfanilate (4-aminobenzenesulfonate) by a novel variation of the β-ketoadipate pathway via 4-sulfocatechol and 3-sulfomuconate. It was previously proposed that the further metabolism of 3-sulfomuconate is catalysed by modified 3-carboxy-cis,cis-muconate-lactonizing enzymes (CMLEs) and that these ‘type 2’ enzymes were different from the conventional CMLEs (‘type 1’) from the protocatechuate pathway in their ability to convert 3-sulfomuconate in addition to 3-carboxy-cis,cis-muconate. In the present study the genes for two CMLEs (pcaB2S1 and pcaB2S2) were cloned from H. intermedia S1 and A. radiobacter S2, respectively. In both strains, these genes were located close to the previously identified genes encoding the 4-sulfocatechol-converting enzymes. The gene products of pcaB2S1 and pcaB2S2 were therefore tentatively identified as type 2 enzymes involved in the metabolism of 3-sulfomuconate. The genes were functionally expressed and the gene products were shown to convert 3-carboxy-cis,cis-muconate and 3-sulfomuconate. 4-Carboxymethylene-4-sulfo-but-2-en-olide (4-sulfomuconolactone) was identified by HPLC-MS as the product, which was enzymically formed from 3-sulfomuconate. His-tagged variants of both CMLEs were purified and compared with the CMLE from the protocatechuate pathway of Pseudomonas putida PRS2000 for the conversion of 3-carboxy-cis,cis-muconate and 3-sulfomuconate. The CMLEs from the 4-sulfocatechol pathway converted 3-sulfomuconate with considerably higher activities than 3-carboxy-cis,cis-muconate. Also the CMLE from P. putida converted 3-sulfomuconate, but this enzyme demonstrated a clear preference for 3-carboxy-cis,cis-muconate as substrate. Thus it was demonstrated that in the 4-sulfocatechol pathway, distinct CMLEs are formed, which are specifically adapted for the preferred conversion of sulfonated substrates.

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Variation in Genes Encoding the Neuroactive Steroid Synthetic Enzymes 5α-Reductase Type 1 and 3α-Reductase Type 2 Is Associated With Alcohol Dependence

Alcoholism Clinical and Experimental Research ◽

10.1111/j.1530-0277.2010.01425.x ◽

2011 ◽

Vol 35 (5) ◽

pp. 946-952 ◽

Cited By ~ 26

Author(s):

Verica Milivojevic ◽

Henry R. Kranzler ◽

Joel Gelernter ◽

Linda Burian ◽

Jonathan Covault

Keyword(s):

Alcohol Dependence ◽

Neuroactive Steroid ◽

Synthetic Enzymes ◽

Genes Encoding

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Differential recognition of ORF2 protein from type 1 and type 2 porcine circoviruses and identification of immunorelevant epitopes

Microbiology ◽

10.1099/0022-1317-81-7-1815 ◽

2000 ◽

Vol 81 (7) ◽

pp. 1815-1824 ◽

Cited By ~ 159

Author(s):

Dominique Mahé ◽

Philippe Blanchard ◽

Catherine Truong ◽

Claire Arnauld ◽

Pierre Le Cann ◽

...

Keyword(s):

Porcine Circovirus ◽

Diagnostic Tools ◽

Wasting Syndrome ◽

Orf2 Protein ◽

Genes Encoding ◽

Porcine Circoviruses ◽

Specific Antigens ◽

Identity Based

Two types of porcine circovirus (PCV) have been isolated and are referred to as PCV1 and PCV2. PCV1 represents an apathogenic virus, whereas PCV2 is associated with post-weaning multisystemic wasting syndrome. The two PCVs are related, since they display about 70% identity based on nucleotide sequences. In order to discriminate between common and type-specific antigens, an immunocytological approach was used following transfections with cloned circovirus DNAs, as well as recombinant proteins expressed by either baculovirus or plasmid vectors. The ORF1-encoded proteins in the two viruses were shown to be antigenically related, whereas the ORF2 proteins were recognized differentially by polyclonal anti-PCV2 antibodies. Furthermore, PEPSCAN analysis performed on overlapping fragments of the genes encoding part of ORF1 and the entire ORF2 and ORF3 led to the identification of five dominant immunoreactive areas, one located on ORF1 and four on ORF2. However, only some ORF2 peptides proved to be immunorelevant epitopes for virus type discrimination. The potential use of ORF2-derived antigens as diagnostic tools is demonstrated.

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Search for Genetic Predictors of Adult Autoimmune Polyendocrine Syndrome in Monozygotic Twins

Clinical Medicine Insights Endocrinology and Diabetes ◽

10.1177/11795514211009796 ◽

2021 ◽

Vol 14 ◽

pp. 117955142110097

Author(s):

Marina Yuryevna Yukina ◽

Anna Aleksandrovna Larina ◽

Evgeny Vitalyevich Vasilyev ◽

Ekaterina Anatolyevna Troshina ◽

Diana Arshaluysovna Dimitrova

Keyword(s):

Monozygotic Twins ◽

Regulatory Proteins ◽

Endocrine Glands ◽

Histocompatibility Complex ◽

Autoimmune Polyendocrine Syndrome ◽

Genetic Predictors ◽

Endocrine Organs ◽

Genes Encoding

Autoimmune polyendocrine syndromes (APS) are a heterogeneous group of diseases characterized by the presence of autoimmune dysfunction of 2 or more endocrine glands and other non-endocrine organs. The components of the syndrome can manifest throughout life: in childhood—APS type 1 (the juvenile type) and in adulthood—APS type 2, 3, and 4 (the adult types). Adult types of APS are more common in clinical practice. It is a polygenic disease associated with abnormalities in genes encoding key regulatory proteins of the major histocompatibility complex (MHC). The search of for candidate genes responsible for mutations in adult APS is continuing. Genetic predisposition is insufficient for the manifestation of the APS of adults, since the penetrance of the disease, even among monozygotic twins, does not approach 100% (30–70%). The article presents the case of isolated Addison’s disease and APS type 2 in monozygotic twins with a revealed compound heterozygosity in the candidate gene VTCN1.

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Yeast acyl-CoA-binding protein: acyl-CoA-binding affinity and effect on intracellular acyl-CoA pool size

Biochemical Journal ◽

10.1042/bj3020479 ◽

1994 ◽

Vol 302 (2) ◽

pp. 479-485 ◽

Cited By ~ 68

Author(s):

J Knudsen ◽

N J Faergeman ◽

H Skøtt ◽

R Hummel ◽

C Børsting ◽

...

Keyword(s):

Amino Acid ◽

Southern Blotting ◽

Binding Protein ◽

Amino Acid Residues ◽

Binding Properties ◽

High Affinity ◽

Genes Encoding ◽

The One

Acyl-CoA-binding protein (ACBP) is a 10 kDa protein characterized in vertebrates. We have isolated two ACBP homologues from the yeast Saccharomyces carlsbergensis, named yeast ACBP types 1 and 2. Both proteins contain 86 amino acid residues and are identical except for four conservative substitutions. In comparison with human ACBP, yeast ACBPs exhibit 48% (type 1) and 49% (type 2) conservation of amino acid residues. The amino acid sequence of S. carlsbergensis ACBP type 1 was found to be identical with the one ACBP present in Saccharomyces cerevisiae. A recombinant form of this protein was expressed in Escherichia coli and S. cerevisiae, purified, and its acyl-CoA-binding properties were characterized by isoelectric focusing and microcalorimetric analyses. The yeast ACBP was found to bind acyl-CoA esters with high affinity (Kd 0.55 x 10(-10) M). Overexpression of yeast ACBP in S. cerevisiae resulted in a significant expansion of the intracellular acyl-CoA pool. Finally, Southern-blotting analysis of the two genes encoding ACBP types 1 and 2 in S. carlsbergensis strongly indicated that this species is a hybrid between S. cerevisiae and Saccharomyces monacensis.

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