scholarly journals Investigation of the binding of Ca2+, Mg2+, Mn2+ and K+ to the vitamin D-dependent Ca2+-binding protein from pig duodenum

1984 ◽  
Vol 219 (1) ◽  
pp. 287-292 ◽  
Author(s):  
D T W Bryant ◽  
P Andrews
Keyword(s):  
Vitamin D ◽  
Binding Protein ◽  
Cation Binding ◽  
Binding Studies ◽  
Binding Properties ◽  
Kd Values ◽  
Significant Difference ◽  
The One ◽  
Fluorimetric Titration ◽  
Flow Dialysis

The cation-binding properties of the vitamin D-dependent Ca2+-binding protein from pig duodenum were investigated, mainly by flow dialysis. The protein bound two Ca2+ ions with high affinity, and Mg2+, Mn2+ and K+ were all bound competitively with Ca2+ at both sites. The sites were distinguished by their different affinities for Mn2+, the one with the higher affinity being designated A (Kd 0.61 +/- 0.02 microM) and the other B (Kd 50 +/- 6 microM). Competitive binding studies allied to fluorimetric titration with Mg2+ showed that site A bound Ca2+, Mg2+ and K+ with Kd values of 4.7 +/- 0.8 nM, 94 +/- 18 microM and 1.6 +/- 0.3 mM respectively, and site B bound the same three cations with Kd values of 6.3 +/- 1.8 nM, 127 +/- 38 microM and 2.1 +/- 0.6 mM. For the binding of these cations, therefore, there was no significant difference between the two sites. In the presence of 1 mM-Mg2+ and 150 mM-K+, both sites bound Ca2+ with an apparent Kd of 0.5 microM. The cation-binding properties were discussed relative to those of parvalbumin, troponin C and the vitamin D-dependent Ca2+-binding protein from chick duodenum.

scholarly journals Polymorphism in Vitamin D-Binding Protein as a Genetic Risk Factor in the Pathogenesis of Endometriosis

2010 ◽  
Vol 31 (6) ◽  
pp. 943-944
Author(s):  
Klaus Faserl ◽  
Georg Golderer ◽  
Leopold Kremser ◽  
Herbert Lindner ◽  
Bettina Sarg ◽  
...  
Keyword(s):  
Vitamin D ◽  
Risk Factor ◽  
Acute Phase Proteins ◽  
Binding Protein ◽  
Medical Center ◽  
Cross Sectional Study ◽  
Vitamin D Binding Protein ◽  
Complement Components ◽  
Cross Sectional ◽  
Significant Difference

Context Previous studies have implicated a deficiency in the inflammatory response in women who develop endometriosis. The specific immunological deficits have not been completely elucidated. Objective Our objective was to identify differences in protein expression in serum that might shed light on the pathophysiology of endometriosis. Design and Setting This cross-sectional study of women undergoing laparoscopy between 2003 and 2005 took place at a university medical center. Patients Patients included consenting women age 18-49 yr undergoing surgery for pain and/or infertility or elective tubal ligation. Women with acute or chronic medical conditions were excluded. Intervention Blood was collected preoperatively. Main Outcome Measure Proteomic analysis of serum was done using two-dimensional difference gel electrophoresis. Results We found 25 protein spots with a significant difference in abundance between women with endometriosis and controls, including acute-phase proteins and complement components. The abundance of vitamin D-binding protein was higher in all endometriosis pools by a factor of approximately 3 compared with the control pool (P < 0.02). Analysis of specific allele products using nano-LC-ESI-MS indicated that it was the GC*2 allele product that was in greater concentration in serum pools, as well as in single validation samples, in women with endometriosis (P = 0.006). In contrast to the GC*1 allele product, which is readily converted to a potent macrophage factor (Gc protein-derived macrophage-activating factor), the GC*2 allele product undergoes practically no such conversion. Conclusions We speculate that the inability to sufficiently activate macrophages’ phagocytotic function in those carrying the GC*2 polymorphism (more prevalent in endometriosis) may allow endometriotic tissues to implant in the peritoneal cavity. Future studies evaluating specific vitamin D-binding protein polymorphisms as a risk factor for endometriosis in larger populations of women are warranted.

scholarly journals Quin 2: the dissociation constants of its Ca2+ and Mg2+ complexes and its use in a fluorimetric method for determining the dissociation of Ca2+-protein complexes

1985 ◽  
Vol 226 (2) ◽  
pp. 613-616 ◽  
Author(s):  
D T W Bryant
Keyword(s):  
Vitamin D ◽  
Good Accuracy ◽  
Dissociation Constants ◽  
Protein Complexes ◽  
Binding Protein ◽  
Fluorimetric Method ◽  
Kd Values ◽  
Spectrophotometric Titrations

Fluorimetric or spectrophotometric titrations with the appropriate cations gave Kd values of 2.9 +/- 0.2 nM and 89 +/- 5 microM respectively for the Ca2+ and Mg2+ complexes of quin 2 at pH 7.5. Mixtures of quin 2 and vitamin D-dependent Ca2+-binding protein from pig duodenum were titrated fluorimetrically with Ca2+ in the absence or presence of Mg2+. These measurements were used with the Kd values of the Ca2+ and Mg2+ complexes of quin 2 to obtain Kd or apparent Kd values for Ca2+-protein complexes ranging from 5 nM to 5 microM with good accuracy.

scholarly journals Polymorphism in Vitamin D-Binding Protein as a Genetic Risk Factor in the Pathogenesis of Endometriosis

Endocrinology ◽  
2010 ◽  
Vol 151 (12) ◽  
pp. 5971-5972
Author(s):  
Klaus Faserl ◽  
Georg Golderer ◽  
Leopold Kremser ◽  
Herbert Lindner ◽  
Bettina Sarg ◽  
...  
Keyword(s):  
Vitamin D ◽  
Risk Factor ◽  
Acute Phase Proteins ◽  
Binding Protein ◽  
Medical Center ◽  
Cross Sectional Study ◽  
Vitamin D Binding Protein ◽  
Complement Components ◽  
Cross Sectional ◽  
Significant Difference

Context: Previous studies have implicated a deficiency in the inflammatory response in women who develop endometriosis. The specific immunological deficits have not been completely elucidated. Objective: Our objective was to identify differences in protein expression in serum that might shed light on the pathophysiology of endometriosis. Design and Setting: This cross-sectional study of women undergoing laparoscopy between 2003 and 2005 took place at a university medical center. Patients: Patients included consenting women age 18-49 yr undergoing surgery for pain and/or infertility or elective tubal ligation. Women with acute or chronic medical conditions were excluded. Intervention: Blood was collected preoperatively. Main Outcome Measure: Proteomic analysis of serum was done using two-dimensional difference gel electrophoresis. Results: We found 25 protein spots with a significant difference in abundance between women with endometriosis and controls, including acute-phase proteins and complement components. The abundance of vitamin D-binding protein was higher in all endometriosis pools by a factor of approximately 3 compared with the control pool (P < 0.02). Analysis of specific allele products using nano-LC-ESI-MS indicated that it was the GC*2 allele product that was in greater concentration in serum pools, as well as in single validation samples, in women with endometriosis (P = 0.006). In contrast to the GC*1 allele product, which is readily converted to a potent macrophage factor (Gc protein-derived macrophage-activating factor), the GC*2 allele product undergoes practically no such conversion. Conclusions: We speculate that the inability to sufficiently activate macrophages’ phagocytotic function in those carrying the GC*2 polymorphism (more prevalent in endometriosis) may allow endometriotic tissues to implant in the peritoneal cavity. Future studies evaluating specific vitamin D-binding protein polymorphisms as a risk factor for endometriosis in larger populations of women are warranted.

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

1994 ◽  
Vol 302 (2) ◽  
pp. 479-485 ◽  
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.

No association of vitamin D binding protein gene polymorphisms with ulcerative colitis in Chinese patients

2019 ◽  
Author(s):  
Fubin Qiu ◽  
Lijuan Zhang ◽  
Jing Wang ◽  
Rui Li ◽  
Linxue Yang
Keyword(s):  
Ulcerative Colitis ◽  
Vitamin D ◽  
Binding Protein ◽  
Case Control ◽  
Chinese Patients ◽  
Vitamin D Binding Protein ◽  
Chinese Han ◽  
Gene Encoding ◽  
Significant Difference

Abstract Object Vitamin D (VD) deficiency has been reported in patients with ulcerative colitis (UC), and polymorphism in the gene encoding the vitamin D binding protein (DBP) can affect the characteristics of DBP, thus affecting the level and function of VD in vivo . Previous studies have rarely reported on the potential relationship between DBP polymorphisms and UC. To investigate the associations between genetic variants in DBP genes and UC susceptibility in the Han Chinese population, in order to discern whether any differences exist between this population and those of other countries.Methods In this case-control study, the genotyping of DBP rs4588 and rs7041 polymorphisms was conducted using polymerase chain reaction (PCR)-ligase detection reactions, and the rs4588 and rs7041 genotypes were detected by PCR-restriction fragment length polymorphism.Results In our case-control cohort, no significant difference was observed in the UC risk for either of the two SNPs (rs4588 and rs7401) in the DBP genes ( P > 0.05). No association between UC susceptibility and the DBP gene haplotypes was found either.Conclusions Our results suggest that the two SNPs (rs4588 and rs7401) in the DBP genes may have no correlation with susceptibility to UC in the Chinese Han population. But interestingly, haplotype GC, which contains the rs4588 and rs7041 variants in the DBP gene, may affect the level of oxidative stress in UC patients, especially the level of MDA.

scholarly journals Investigation of the forms of pig duodenal Ca2+-binding protein produced by limited tryptic hydrolysis

1986 ◽  
Vol 237 (3) ◽  
pp. 781-787 ◽  
Author(s):  
D T W Bryant ◽  
S Critch
Keyword(s):  
Binding Site ◽  
Binding Sites ◽  
Binding Protein ◽  
Native Protein ◽  
Binding Studies ◽  
High Affinity ◽  
C Terminus ◽  
Kd Values ◽  
T1 And T2 ◽  
Tryptic Hydrolysis

Vitamin D-dependent Ca2+-binding protein from pig duodenum was hydrolysed with trypsin in the presence of Ca2+ and two products were obtained: T1, which differed from the native protein by loss of Ac-Ser-Ala-Gln-Lys from the N-terminus and Ile-Ser-Gln-OH from the C-terminus, and T2, which differed from T1 by loss of a C-terminal lysine. The hydrolysis inactivated one of the two high-affinity Ca2+-binding sites on the native protein, and the remaining site was stable in T1 but labile in T2 when the proteins were Ca2+-free. Binding studies showed that T1 had Kd values of 2.8 +/- 0.1 nM, 57 +/- 13 microM and 0.8 +/- 0.3 microM for Ca2+, Mg2+ and Mn2+ respectively, and T2 had Kd 2.2 +/- 0.3 nM for Ca2+. The affinity for Mn2+, together with the other Kd values, identified the site on T1 as the site on the native protein previously found to have Kd 0.6 microM for Mn2+, rather than one with Kd 50 microM for Mn2+. In contrast with both the native protein and another form of the protein with a single Ca2+-binding site, the intrinsic fluorescence of T1 and T2 was little affected by the addition of Ca2+. It was concluded that the active binding site in T1 and T2, and also the site in the native protein with the higher affinity for Mn2+, was probably in the C-terminal half of the molecule.

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