Characterization of a Truncated Form of Recombinant Porcine Growth Hormone Generated in Vitro During Solubilization of Inclusion Bodies

1993 ◽  
Vol 4 (2) ◽  
pp. 164-175 ◽  
Author(s):  
N.K. Puri ◽  
M. Cardamone ◽  
E. Crivelli ◽  
J.C. Traeger
Keyword(s):  
Growth Hormone ◽  
Inclusion Bodies ◽  
Truncated Form ◽  
Solubilization Of Inclusion Bodies

scholarly journals Characterization of a Dye-Decolorizing Peroxidase from Irpex lacteus Expressed in Escherichia coli: An Enzyme with Wide Substrate Specificity Able to Transform Lignosulfonates

2021 ◽  
Vol 7 (5) ◽  
pp. 325
Author(s):  
Laura Isabel de de Eugenio ◽  
Rosa Peces-Pérez ◽  
Dolores Linde ◽  
Alicia Prieto ◽  
Jorge Barriuso ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Inclusion Bodies ◽  
Veratryl Alcohol ◽  
Dye Decolorization ◽  
Irpex Lacteus ◽  
Type D ◽  
Lignin Peroxidases

A dye-decolorizing peroxidase (DyP) from Irpex lacteus was cloned and heterologously expressed as inclusion bodies in Escherichia coli. The protein was purified in one chromatographic step after its in vitro activation. It was active on ABTS, 2,6-dimethoxyphenol (DMP), and anthraquinoid and azo dyes as reported for other fungal DyPs, but it was also able to oxidize Mn2+ (as manganese peroxidases and versatile peroxidases) and veratryl alcohol (VA) (as lignin peroxidases and versatile peroxidases). This corroborated that I. lacteus DyPs are the only enzymes able to oxidize high redox potential dyes, VA and Mn+2. Phylogenetic analysis grouped this enzyme with other type D-DyPs from basidiomycetes. In addition to its interest for dye decolorization, the results of the transformation of softwood and hardwood lignosulfonates suggest a putative biological role of this enzyme in the degradation of phenolic lignin.

Characterization of the somatostatin-like immunoreactivity extracted from an adrenal medullary tumour

1982 ◽  
Vol 2 (3) ◽  
pp. 147-154 ◽  
Author(s):  
R. Corder ◽  
J. E. C. Sykes ◽  
P. J. Lowry
Keyword(s):  
Growth Hormone ◽  
Anterior Pituitary ◽  
Gel Filtration ◽  
Hormone Release ◽  
Growth Hormone Release ◽  
Identical Composition ◽  
Salt Fractionation ◽  
Somatostatin 14

Significant amounts of somatostatin-like immunor reactivity (SLI) were detected in the extract of a human catecholamine-secreting adrenal medullary tumour. After salt fractionation and reconstitution the major portion of SLI was purified by gel filtration and two HPLC steps; in all three systems it eluted in the position of somatostatin-14. The purified somatostatin-like peptide inhibited, in a dose-related manner, growth hormone release from stimulated perfused rat anterior pituitary ceils in vitro. Amino acid analysis showed the purified peptide to have an identical composition to somatostatin found in other species.

scholarly journals Functional characterization of canine wild type glucocorticoid receptor and an insertional mutation in a dog

2019 ◽  
Vol 15 (1) ◽  
Author(s):  
Kosei Yamanaka ◽  
Masaru Okuda ◽  
Takuya Mizuno
Keyword(s):  
Glucocorticoid Receptor ◽  
Cushing Syndrome ◽  
Functional Characterization ◽  
Wild Type ◽  
Key Factors ◽  
Truncated Form ◽  
Glucocorticoid Sensitivity ◽  
Almost All

Abstract Background Glucocorticoids, among the most widely utilized drugs in veterinary medicine, are employed to treat a wide variety of diseases; however, their use often induces adverse events in dogs. The efficacy of glucocorticoids usually depends on dosage, although differences in sensitivity to glucocorticoids in individual animals have been reported. Glucocorticoids bind to the cytoplasmic glucocorticoid receptor (GR), which is expressed in almost all cells. These receptors are key factors in determining individual sensitivity to glucocorticoids. This study examined individual differences in glucocorticoid sensitivity in dogs, focusing on reactivity of the GR to prednisolone. Results We first molecularly cloned the GR gene from a healthy dog. We discovered a mutant GR in a dog suspected to have iatrogenic Cushing syndrome. The mutant GR had extra nucleotides between exons 6 and 7, resulting in a truncated form of GR that was 98 amino acids shorter than the wild-type dog GR. The truncated GR exhibited very low reactivity to prednisolone, irrespective of concentration. Conclusions We have identified the truncated form of canine GR in a dog with iatrogenic Cushing syndrome. This truncated form showed the very less sensitivity to glucocorticoid in vitro, unfortunately, we could not elucidate its clinical significance. However, our data is a first report about the function of canine GR, and will facilitate the analysis of canine glucocorticoid sensitivity.

scholarly journals Detection and characterization of proteins encoded by the second ORF of the M2 gene of pneumoviruses

1999 ◽  
Vol 80 (8) ◽  
pp. 2011-2016 ◽  
Author(s):  
G. Ahmadian ◽  
P. Chambers ◽  
A. J. Easton
Keyword(s):  
Inclusion Bodies ◽  
Virus Protein ◽  
Sequence Identity ◽  
Avian Pneumovirus ◽  
Orf2 Protein ◽  
Monospecific Antisera ◽  
Gst Fusion Proteins

The nucleotide sequence of the M2 gene of pneumonia virus of mice (PVM) was determined. The sequence showed that the gene encoded a protein of 176 amino acids with a predicted molecular mass of 20165 Da from a major ORF, which is smaller than the equivalent proteins encoded by human, bovine and ovine respiratory syncytial (RS) viruses. The PVM M2 protein is conserved, having 41% similarity to the equivalent human RS virus protein. In common with the M2 genes of the RS viruses and avian pneumovirus (APV), the PVM mRNA also contained a second ORF (ORF2) that partially overlaps the first ORF and which is capable of encoding a 98 residue polypeptide. No significant sequence identity could be detected between the putative M2 ORF2 proteins of PVM, APV and the RS viruses. The expression of the M2 ORF2 proteins of the pneumoviruses was investigated by using monospecific antisera raised against GST fusion proteins. Western blot analysis demonstrated the presence of polypeptides encoded by M2 ORF2 of PVM and RS virus corresponding with those predicted by in vitro translation studies, but this was not the case for APV. The PVM polypeptide was present as three distinct products in vivo. The PVM and RS virus polypeptides were also detected in cells by immunofluorescence, which showed that both were present in the cytoplasm with a degree of localization in inclusion bodies. No APV M2 ORF2 protein could be detected in vivo. The RS virus M2 ORF2 polypeptide was shown to accumulate during infection and the potential implications of this are discussed.

Recombinant murine growth hormone from E. coli inclusion bodies: Expression, high-pressure solubilization and refolding, and characterization of activity and structure

2010 ◽  
Vol 26 (3) ◽  
pp. 743-749 ◽  
Author(s):  
Amber Haynes Fradkin ◽  
Carl S. Boand ◽  
Stephen P. Eisenberg ◽  
Mary S. Rosendahl ◽  
Theodore W. Randolph
Keyword(s):  
Growth Hormone ◽  
High Pressure ◽  
Inclusion Bodies ◽  
E Coli

scholarly journals In Vitro and in Vivo Characterization of MOD-4023, a Long-Acting Carboxy-Terminal Peptide (CTP)-Modified Human Growth Hormone

2016 ◽  
Vol 13 (2) ◽  
pp. 631-639 ◽  
Author(s):  
Oren Hershkovitz ◽  
Ahuva Bar-Ilan ◽  
Rachel Guy ◽  
Yana Felikman ◽  
Laura Moschcovich ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Human Growth Hormone ◽  
Human Growth ◽  
Long Acting ◽  
Carboxy Terminal ◽  
In Vivo Characterization

Characterization of a Novel POU1F1 Mutation Identified on Screening 160 Growth Hormone Deficiency Patients

2019 ◽  
Vol 51 (04) ◽  
pp. 248-255
Author(s):  
Shweta Birla ◽  
P Vijayakumar ◽  
Shilpi Sehgal ◽  
Shinjini Bhatnagar ◽  
Kshetrapal Pallavi ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Growth Hormone Deficiency ◽  
Functional Characterization ◽  
Failure To Thrive ◽  
Site Directed Mutagenesis ◽  
Hormone Deficiency ◽  
Pituitary Hormone ◽  
Severe Short Stature

AbstractThe objective of the study is the functional characterization of a novel POU1F1 c.605delC mutation in combined pituitary hormone deficiency (CPHD) and to report the clinical and genetic details of 160 growth hormone deficiency patients. Screening of GH1, GHRHR, POU1F1, PROP1, and HESX1 genes by Sanger sequencing was carried out in 160 trios and 100 controls followed by characterization of the POU1F1 c.605delC mutation by expression studies including site directed mutagenesis, co-transfection, protein degradation, and luciferase assays to compare the wild type and mutant POU1F1. In vitro studies showed that the POU1F1 c.605delC mutation codes for a truncated protein with reduced transactivation capacity on its downstream effectors, viz., growth hormone (GH) and prolactin (PRL) causing severe CPHD. Experiments using different protease inhibitors reveal rescue of the protein upon blockage of the lysosomal pathway that might be useful in novel drug designing using targeted approach thereby maintaining the milieu and preventing/delaying the disease. The study provides an insight into the disease causing mechanism of POU1F1 c.605delC mutation identified in a CPHD child with severe short stature and failure to thrive. It also shows mutation effect on the expression, function and turnover of protein and highlights mechanistic details by which these potent regulators may operate.

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