scholarly journals Introduction of histidine residues into avidin subunit interfaces allows pH-dependent regulation of quaternary structure and biotin binding

FEBS Letters ◽  
2003 ◽  
Vol 555 (3) ◽  
pp. 449-454 ◽  
Author(s):  
Henri R Nordlund ◽  
Vesa P Hytönen ◽  
Olli H Laitinen ◽  
Sanna T.H Uotila ◽  
Einari A Niskanen ◽  
...  
Keyword(s):  
Quaternary Structure ◽  
Histidine Residues ◽  
Biotin Binding ◽  
Ph Dependent

scholarly journals Generating a High Valency Biotin Binder by Selecting Uniform Protein Assemblies via Crystallization

Crystals ◽  
2019 ◽  
Vol 9 (7) ◽  
pp. 353
Author(s):  
Orly Avraham ◽  
Yael Levi-Kalisman ◽  
Oded Livnah
Keyword(s):  
Quaternary Structure ◽  
Disulfide Bridge ◽  
Symmetry Operation ◽  
Added Value ◽  
Purification Process ◽  
Protein Assemblies ◽  
High Affinity ◽  
Covalent Crosslinking ◽  
Crystallographic Symmetry ◽  
Biotin Binding

Crystallization is a common practice in the purification process in small molecule synthesis while selecting the wanted product. For proteins it is rarely applied due to the methodological predicaments in obtaining crystals. Our observation of the stabilized octamers in the crystal structure of hoefavidin, a novel dimeric member of the avidin family, led to the notion of developing a novel biotechnological tool via covalent crosslinking. The avidin–biotin system has been exploited for decades utilizing the ultra-high affinity between avidin and biotin as a basis for numerous applications. Optimizing the system led to the discovery of a novel group of dimeric avidins including hoefavidin. Hoefavidin has a dynamic quaternary structure, where a dimer is the basis for generating the octamer via crystallographic symmetry operation. Upon biotin binding in solution hoefavidin dissociates solely into dimers. In order to stabilize the octamer, we designed the P61C mutant to form a disulfide bridge stabilizing the octamer and preventing dissociation upon biotin binding. The process of selecting P61C hoefavidin uniform octamers includes crystallization followed by dissolving the crystals. The P61C modified hoefavidin octamer can have a substantial added value to the various biotechnological applications and advances of the biotin based high affinity systems.

scholarly journals Phylogenetic and Functional Analysis of Histidine Residues Essential for pH-dependent Multimerization of von Willebrand Factor

2011 ◽  
Vol 286 (29) ◽  
pp. 25763-25769 ◽  
Author(s):  
Luke T. Dang ◽  
Angie R. Purvis ◽  
Ren-Huai Huang ◽  
Lisa A. Westfield ◽  
J. Evan Sadler
Keyword(s):  
Functional Analysis ◽  
Von Willebrand Factor ◽  
Histidine Residues ◽  
Ph Dependent ◽  
Von Willebrand ◽  
Willebrand Factor

scholarly journals Mutation of the important Tyr-33 residue of chicken avidin: functional and structural consequences

2003 ◽  
Vol 369 (2) ◽  
pp. 249-254 ◽  
Author(s):  
Ari T. MARTTILA ◽  
Vesa P. HYTÖNEN ◽  
Olli H. LAITINEN ◽  
Edward A. BAYER ◽  
Meir WILCHEK ◽  
...  
Keyword(s):  
Ligand Binding ◽  
Quaternary Structure ◽  
Affinity Purification ◽  
Ph Dependence ◽  
Tight Binding ◽  
Carbonyl Oxygen ◽  
Binding Characteristics ◽  
Biotin Binding ◽  
High Ligand

The strong interaction between avidin and biotin is so tight (dissociation constant 10-15M) that conditions usually sufficient for protein denaturing fail to dislodge biotin from the avidin—biotin complex. This kind of irreversible binding hinders the use of avidin in applications such as affinity purification or protein immobilization. To address this concern, we have constructed a series of mutants of the strategically positioned Tyr-33 in order to study the role of this residue in biotin binding, and to create avidin variants with more reversible ligand-binding properties. Unexpectedly, an avidin mutant in which Tyr-33 was replaced with phenylalanine (Avm-Y33F) displayed similar biotin-binding characteristics to the native avidin, indicating that the hydrogen bond formed between the hydroxy group of Tyr-33 and the carbonyl oxygen of biotin is not as important for the tight binding of biotin as previously suggested. In terms of the reversibility of biotin binding, Avm-Y33H was the most successful substitution constructed in this study. Interestingly, the binding of this mutant exhibited clear pH-dependence, since at neutral pH it bound to the biotin surface in an irreversible fashion, whereas, at pH9, 50% of the bound protein could be released with free biotin. Furthermore, although Tyr-33 is located relatively distant from the monomer—monomer interfaces, the mutagenesis of this residue also weakened the quaternary structure of avidin, indicating that the high ligand binding and the high stability of avidin have evolved together and it is difficult to modify one without affecting the other.

scholarly journals Protonation of Individual Histidine Residues Is Not Required for the pH-Dependent Entry of West Nile Virus: Evaluation of the “Histidine Switch” Hypothesis

2009 ◽  
Vol 83 (23) ◽  
pp. 12631-12635 ◽  
Author(s):  
Steevenson Nelson ◽  
Subhajit Poddar ◽  
Tsai-Yu Lin ◽  
Theodore C. Pierson
Keyword(s):  
Life Cycle ◽  
West Nile Virus ◽  
Fusion Proteins ◽  
Viral Fusion ◽  
Ph Sensing ◽  
West Nile ◽  
Histidine Residues ◽  
E Protein ◽  
Viral Fusion Proteins ◽  
Ph Dependent

ABSTRACT Histidine residues have been hypothesized to function as sensors of environmental pH that can trigger the activity of viral fusion proteins. We investigated a requirement for histidine residues in the envelope (E) protein of West Nile virus during pH-dependent entry into cells. Each histidine was individually replaced with a nonionizable amino acid and tested functionally. In each instance, mutants capable of orchestrating pH-dependent infection were identified. These results do not support a requirement for any single histidine as a pH-sensing “switch,” and they suggest that additional features of the E protein are involved in triggering pH-dependent steps in the flavivirus life cycle.

scholarly journals Large-Scale, pH-Dependent, Quaternary Structure Changes in an RNA Virus Capsid Are Reversible in the Absence of Subunit Autoproteolysis

2002 ◽  
Vol 76 (19) ◽  
pp. 9972-9980 ◽  
Author(s):  
Derek J. Taylor ◽  
Neel K. Krishna ◽  
Mary A. Canady ◽  
Anette Schneemann ◽  
John E. Johnson
Keyword(s):  
Coat Protein ◽  
Conformational Change ◽  
Conformational Changes ◽  
Large Scale ◽  
Quaternary Structure ◽  
Rna Virus ◽  
Expression System ◽  
Recombinant Baculovirus ◽  
Baculovirus Expression System ◽  
Ph Dependent

ABSTRACT The assembly and maturation of the coat protein of a T=4, nonenveloped, single-stranded RNA virus, Nudaurelia capensis ω virus (NωV), was examined by using a recombinant baculovirus expression system. At pH 7.6, the coat protein assembles into a stable particle called the procapsid, which is 450 Å in diameter and porous. Lowering the pH to 5.0 leads to a concerted reorganization of the subunits into a 410-Å-diameter particle called the capsid, which has no obvious pores. This conformational change is rapid but reversible until slow, autoproteolytic cleavage occurs in at least 15% of the subunits at the lower pH. In this report, we show that expression of subunits with replacement of Asn-570, which is at the cleavage site, with Thr results in assembly of particles with expected morphology but that are cleavage defective. The conformational change from procapsid to capsid is reversible in N570T mutant virus-like particles, in contrast to wild-type particles, which are locked into the capsid conformation after cleavage of the coat protein. The reexpanded procapsids display slightly different properties than the original procapsid, suggesting hysteretic effects. Because of the stability of the procapsid under near-neutral conditions and the reversible properties of the cleavage-defective mutant, NωV provides an excellent model for the study of pH-induced conformational changes in macromolecular assemblies. Here, we identify the relationship between cleavage and the conformational change and propose a pH-dependent helix-coil transition that may be responsible for the structural rearrangement in NωV.

A Monoclonal Antibody to Avidin Dissociates Quaternary Structure and Curtails Biotin Binding to Avidin and Streptavidin

1997 ◽  
Vol 344 (2) ◽  
pp. 281-288 ◽  
Author(s):  
N. Subramanian ◽  
Sarada Subramanian ◽  
Anjali A. Karande ◽  
P.Radhakantha Adiga
Keyword(s):  
Monoclonal Antibody ◽  
Quaternary Structure ◽  
Biotin Binding

Involvement of Histidine Residues in the pH-Dependent β-Galactoside Binding Activity of Human Galectin-1

Biochemistry ◽  
2013 ◽  
Vol 52 (13) ◽  
pp. 2371-2380 ◽  
Author(s):  
Hirotsugu Hiramatsu ◽  
Katsuyuki Takeuchi ◽  
Hideo Takeuchi
Keyword(s):  
Binding Activity ◽  
Histidine Residues ◽  
Ph Dependent

scholarly journals Regulation of skeletal-muscle AMP deaminase: involvement of histidine residues in the pH-dependent inhibition of the rabbit enzyme by ATP

1995 ◽  
Vol 309 (3) ◽  
pp. 845-852 ◽  
Author(s):  
M Ranieri-Raggi ◽  
F Ronca ◽  
A Sabbatini ◽  
A Raggi
Keyword(s):  
Skeletal Muscle ◽  
Regulatory Mechanism ◽  
Limited Proteolysis ◽  
Acidic Ph ◽  
Amp Deaminase ◽  
Histidine Residues ◽  
Dependent Inhibition ◽  
Inhibitory Site ◽  
Ph Dependent ◽  
Positively Charged

Reaction of rabbit skeletal-muscle AMP deaminase with a low molar excess of diethyl pyrocarbonate results in conversion of the enzyme into a species with one or two carbethoxylated histidine residues per subunit that retains sensitivity to ATP at pH 7.1 but, unlike the native enzyme, it is not sensitive to regulation by ATP at pH 6.5. This effect mimics that exerted on the enzyme by limited proteolysis with trypsin, which removes the 95-residue N-terminal region from the 80 kDa enzyme subunit. These observations suggest involvement of some histidine residues localized in the region HHEMQAHILH (residues 51-60) in the regulatory mechanism which stabilizes the binding of ATP to its inhibitory site at acidic pH. Carbethoxylation of two histidine residues per subunit abolishes the inhibition by ATP of the proteolysed enzyme at pH 7.1, suggesting the obligatory participation of a second class of histidine residues, localized in the 70 kDa subunit core, in the mechanism of the pH-dependent inhibition of the enzyme by ATP. At a slightly acidic pH, these histidine residues would be positively charged, resulting in a desensitized form of the enzyme similar to that obtained with the carbethoxylation reaction.

scholarly journals Histidine Residues Are Responsible for Bidirectional Effects of Zinc on Acid-Sensing Ion Channel 1a/3 Heteromeric Channels

Biomolecules ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1264
Author(s):  
Qian Jiang ◽  
Andrew M. Peterson ◽  
Yuyang Chu ◽  
Xiaolan Yao ◽  
Xiang-ming Zha ◽  
...  
Keyword(s):  
Ion Channel ◽  
Ph Dependence ◽  
Critical Role ◽  
Extracellular Domain ◽  
Significant Inhibition ◽  
Patch Clamp Technique ◽  
Histidine Residues ◽  
Whole Cell Patch Clamp ◽  
Ph Dependent ◽  
Bidirectional Effects

Acid-sensing ion channel (ASIC) subunits 1a and 3 are highly expressed in central and peripheral sensory neurons, respectively. Endogenous biomolecule zinc plays a critical role in physiological and pathophysiological conditions. Here, we found that currents recorded from heterologously expressed ASIC1a/3 channels using the whole-cell patch-clamp technique were regulated by zinc with dual effects. Co-application of zinc dose-dependently potentiated both peak amplitude and the sustained component of heteromeric ASIC1a/3 currents; pretreatment with zinc between 3 to 100 µM exerted the same potentiation as co-application. However, pretreatment with zinc induced a significant inhibition of heteromeric ASIC1a/3 channels when zinc concentrations were over 250 µM. The potentiation of heteromeric ASIC1a/3 channels by zinc was pH dependent, as zinc shifted the pH dependence of ASIC1a/3 currents from a pH50 of 6.54 to 6.77; whereas the inhibition of ASIC1a/3 currents by zinc was also pH dependent. Furthermore, we systematically mutated histidine residues in the extracellular domain of ASIC1a or ASIC3 and found that histidine residues 72 and 73 in both ASIC1a and ASIC3, and histidine residue 83 in the ASIC3 were responsible for bidirectional effects on heteromeric ASIC1a/3 channels by zinc. These findings suggest that histidine residues in the extracellular domain of heteromeric ASIC1a/3 channels are critical for zinc-mediated effects.

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