Catecholamine Binding Protein: Binding of Tritium to a Specific Protein Fraction of Human Plasma following in vitro Incubation with Tritiated Noradrenaline

Nature ◽  
1966 ◽  
Vol 212 (5067) ◽  
pp. 1270-1271 ◽  
Author(s):  
BERNARD L. MIRKIN ◽  
DAVID M. BROWN ◽  
ROBERT A. ULSTROM
Keyword(s):  
Protein Binding ◽  
Human Plasma ◽  
Protein Fraction ◽  
Binding Protein ◽  
Specific Protein ◽  
In Vitro Incubation

EVALUATION OF TISSUE STEROID BINDING IN VITRO

1971 ◽  
Vol 68 (1_Suppl) ◽  
pp. S223-S246 ◽  
Author(s):  
C. R. Wira ◽  
H. Rochefort ◽  
E. E. Baulieu
Keyword(s):  
Protein Binding ◽  
Binding Sites ◽  
Experimental System ◽  
Specific Protein ◽  
Coupling Mechanism ◽  
Target Tissue ◽  
Protein Binding Sites ◽  
Definition Of ◽  
Hormone Specificity

ABSTRACT The definition of a RECEPTOR* in terms of a receptive site, an executive site and a coupling mechanism, is followed by a general consideration of four binding criteria, which include hormone specificity, tissue specificity, high affinity and saturation, essential for distinguishing between specific and nonspecific binding. Experimental approaches are proposed for choosing an experimental system (either organized or soluble) and detecting the presence of protein binding sites. Techniques are then presented for evaluating the specific protein binding sites (receptors) in terms of the four criteria. This is followed by a brief consideration of how receptors may be located in cells and characterized when extracted. Finally various examples of oestrogen, androgen, progestagen, glucocorticoid and mineralocorticoid binding to their respective target tissues are presented, to illustrate how researchers have identified specific corticoid and mineralocorticoid binding in their respective target tissue receptors.

scholarly journals hnRNP K Supports High-Amplitude D Site-Binding Protein mRNA (Dbp mRNA) Oscillation To Sustain Circadian Rhythms

2020 ◽  
Vol 40 (6) ◽  
Author(s):  
Paul Kwangho Kwon ◽  
Kyung-Ha Lee ◽  
Ji-hyung Kim ◽  
Sookil Tae ◽  
Seokjin Ham ◽  
...  
Keyword(s):  
Clock Genes ◽  
Binding Protein ◽  
Underlying Mechanism ◽  
High Amplitude ◽  
Specific Protein ◽  
Proximal Promoter ◽  
Hnrnp K ◽  
Protein Levels ◽  
Site Binding

ABSTRACT Circadian gene expression is defined by the gene-specific phase and amplitude of daily oscillations in mRNA and protein levels. D site-binding protein mRNA (Dbp mRNA) shows high-amplitude oscillation; however, the underlying mechanism remains elusive. Here, we demonstrate that heterogeneous nuclear ribonucleoprotein K (hnRNP K) is a key regulator that activates Dbp transcription via the poly(C) motif within its proximal promoter. Biochemical analyses identified hnRNP K as a specific protein that directly associates with the poly(C) motif in vitro. Interestingly, we further confirmed the rhythmic binding of endogenous hnRNP K within the Dbp promoter through chromatin immunoprecipitation as well as the cycling expression of hnRNP K. Finally, knockdown of hnRNP K decreased mRNA oscillation in both Dbp and Dbp-dependent clock genes. Taken together, our results show rhythmic protein expression of hnRNP K and provide new insights into its function as a transcriptional amplifier of Dbp.

scholarly journals Cooperative Assembly of an hnRNP Complex Induced by a Tissue-Specific Homolog of Polypyrimidine Tract Binding Protein

2000 ◽  
Vol 20 (20) ◽  
pp. 7463-7479 ◽  
Author(s):  
Vadim Markovtsov ◽  
Julia M. Nikolic ◽  
Joseph A. Goldman ◽  
Christoph W. Turck ◽  
Min-Yuan Chou ◽  
...  
Keyword(s):  
Binding Protein ◽  
Regulatory Protein ◽  
Regulatory Elements ◽  
Specific Protein ◽  
Regulatory Sequence ◽  
Polypyrimidine Tract ◽  
Tissue Specific ◽  
Polypyrimidine Tract Binding ◽  
Polypyrimidine Tract Binding Protein

ABSTRACT Splicing of the c-src N1 exon in neuronal cells depends in part on an intronic cluster of RNA regulatory elements called the downstream control sequence (DCS). Using site-specific cross-linking, RNA gel shift, and DCS RNA affinity chromatography assays, we characterized the binding of several proteins to specific sites along the DCS RNA. Heterogeneous nuclear ribonucleoprotein (hnRNP) H, polypyrimidine tract binding protein (PTB), and KH-type splicing-regulatory protein (KSRP) each bind to distinct elements within this sequence. We also identified a new 60-kDa tissue-specific protein that binds to the CUCUCU splicing repressor element of the DCS RNA. This protein was purified, partially sequenced, and cloned. The new protein (neurally enriched homolog of PTB [nPTB]) is highly homologous to PTB. Unlike PTB, nPTB is enriched in the brain and in some neural cell lines. Although similar in sequence, nPTB and PTB show significant differences in their properties. nPTB binds more stably to the DCS RNA than PTB does but is a weaker repressor of splicing in vitro. nPTB also greatly enhances the binding of two other proteins, hnRNP H and KSRP, to the DCS RNA. These experiments identify specific cooperative interactions between the proteins that assemble onto an intricate splicing-regulatory sequence and show how this hnRNP assembly is altered in different cell types by incorporating different but highly related proteins.

Synthesis, crystal structure of novel unsymmetrical heterocyclic Schiff base Ni(II)/V(IV) complexes: Investigation of DNA binding, protein binding and in vitro cytotoxic activity

2019 ◽  
Vol 488 ◽  
pp. 182-194 ◽  
Author(s):  
Maryam Sedighipoor ◽  
Ali Hossein Kianfar ◽  
Gholamhossein Mohammadnezhad ◽  
Helmar Görls ◽  
Winfried Plass ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Schiff Base ◽  
Dna Binding ◽  
Protein Binding ◽  
Cytotoxic Activity ◽  
Binding Protein ◽  
Dna Binding Protein

scholarly journals Cell-cycle-specific transcription termination within the human histone H3.3 gene is correlated with specific protein–DNA interactions

1997 ◽  
Vol 69 (2) ◽  
pp. 101-110 ◽  
Author(s):  
ALAN TAYLOR ◽  
LIQUN ZHANG ◽  
JOHN HERRMANN ◽  
BEI WU ◽  
LARRY KEDES ◽  
...  
Keyword(s):  
Protein Binding ◽  
Rna Polymerase Ii ◽  
Transcription Termination ◽  
Transcription Elongation ◽  
Specific Protein ◽  
Binding Activity ◽  
Mobility Shift ◽  
Nuclear Extracts

In vitro studies using highly purified calf thymus RNA polymerase II and a fragment spanning the first intron of H3.3 as template DNA have demonstrated the existence of a strong transcription termination site consisting of thymidine stretches. In this study, nuclear run-on experiments have been performed to assess the extent to which transcription elongation is blocked in vivo using DNA probes corresponding to regions 5′ and 3′ of the in vitro termination sites. These studies suggest that H3.3 expression is stimulated following the inhibition of DNA synthesis through the elimination of the transcription elongation block. Interestingly, both the in vivo and in vitro experiments have revealed that the transcriptional block/termination sites are positioned immediately downstream of a 73 bp region that has been over 90% conserved between the chicken and human H3.3 genes. The extreme conservation of this intronic region suggests a possible role in maintaining cis-acting function. Electrophoretic mobility shift experiments show that HeLa cell nuclear extracts contain protein factors that bind specifically to the region of transcription elongation block. Furthermore, we demonstrate a correlation between the protein binding activity and the transcriptional block in cells that have been either arrested at the initiation of S phase or were replication-interrupted by hydroxyurea. DNA footprinting experiments indicate that the region of protein binding is at the 3′ end of the conserved region and overlaps with one of the three in-vitro-mapped termination sites.

Role of high-affinity folate-binding protein in the plasma distribution of tetrahydrofolate in pigs

1996 ◽  
Vol 270 (1) ◽  
pp. R105-R110 ◽  
Author(s):  
K. Sasaki ◽  
M. Natsuhori ◽  
M. Shimoda ◽  
Y. Saima ◽  
E. Kokue
Keyword(s):  
Protein Binding ◽  
Binding Capacity ◽  
Binding Protein ◽  
Folate Binding Protein ◽  
High Affinity ◽  
Plasma Distribution ◽  
The Stability ◽  
Plasma Ultrafiltrate

Stability and protein-binding properties of tetrahydrofolate (THF) in pig plasma were studied in vitro. THF in plasma was stable for more than 120 min when it existed in a bound form, whereas THF both in plasma ultrafiltrate and in plasma ultrafiltrate plus porcine albumin was degraded rapidly and disappeared soon after its addition. These results suggest that high-affinity folate-binding protein (HFBP) is related to the stability of THF. THF-protein binding kinetic analysis showed that porcine plasma had HFBP and low-affinity binding protein (albumin) for THF. Dissociation constant and maximal binding capacity of HFBP were calculated to be 0.4 and 70 nM, respectively, indicating that > 98% of endogenous plasma THF existed in bound form with HFBP. Porcine albumin was not essentially a protein that binds and protects endogenous THF from degradation. We conclude that most endogenous THF binds to HFBP and only the unbound form of THF is rapidly degraded in pig plasma. HFBP protects THF from degradation and allows THF to exist stably in pig plasma. In addition, HFBP may govern the species specificity of plasma folate distribution in pigs.

scholarly journals In Vitro Bactericidal Activity of Iclaprim in Human Plasma

2009 ◽  
Vol 53 (10) ◽  
pp. 4542-4544 ◽  
Author(s):  
Heike Laue ◽  
Tiziana Valensise ◽  
Aurélie Seguin ◽  
Sergio Lociuro ◽  
Khalid Islam ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Protein Binding ◽  
Human Plasma ◽  
Bactericidal Activity ◽  
The Novel

ABSTRACT This study evaluated the effect of human plasma on the in vitro bactericidal activity of the novel diaminopyrimidine iclaprim against methicillin (meticillin)-susceptible and -resistant Staphylococcus aureus strains. MICs and minimal bactericidal concentrations (MBCs) of iclaprim, with ∼93% protein binding, were similar in the absence and in the presence of 50% human plasma; MICs and MBCs ranged from 0.06 to 0.125 μg/ml. Furthermore, the activity of iclaprim was not affected by plasma, with ≥99.9% reduction in CFU after 5.0 to 7.6 h.

Reliability of Plasma Cortisol Determination by a Simple Competitive Protein Binding Method

1976 ◽  
Vol 13 (1-6) ◽  
pp. 371-378 ◽  
Author(s):  
G. C. Cashmore ◽  
J. D. Few ◽  
Christine H. Fowles
Keyword(s):  
Protein Binding ◽  
Human Plasma ◽  
Cortisol Level ◽  
Plasma Cortisol ◽  
Binding Protein ◽  
Venous Occlusion ◽  
Plasma Cortisol Level ◽  
Short Term ◽  
Term Variation ◽  
Competitive Protein Binding

The method investigated used [3H] cortisol as the ligand, diluted human plasma as the binding protein, and florisil to separate free and bound cortisol. The precision and specificity were found to be adequate for routine use. The effects of such factors as delay in separation of plasma, storage of plasma, provenance of the binding protein, venepuncture, and venous occlusion have been studied. The extent and significance of short term, inter-daily, and longer term variation in plasma cortisol level are discussed.

Sign in / Sign up

Export Citation Format

Share Document