[15] Determination of rate and equilibrium binding constants for macromolecular interactions by surface plasmon resonance

O’Shannessy, Daniel J.; Brigham-Burke, Michael; Soneson, K. Karl; Hensley, Preston; Brooks, Ian

doi:10.1016/s0076-6879(94)40054-7

Cited by 130 publications

(138 citation statements)

References 17 publications

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“…The dissociation rate constant, k d , was determined by fitting data to the equation R =R 0 e − kd(t − t0) where R 0 is the response level at the beginning of the dissociation phase. This model, which has recently been applied to describe adaptor-cytoplasmic domain interaction (46), is described in more detail elsewhere (47,48). It should be noted that these models allow the determination of rate constants without reaching equilibrium during the experimental cycle.…”

Section: Determination Of Kinetic Rate Constantsmentioning

confidence: 99%

The Simian Immunodeficiency Virus Envelope Glycoprotein Contains Multiple Signals that Regulate its Cell Surface Expression and Endocytosis

Bowers

Pelchen–Matthews

Höning

et al. 2000

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The cell surface expression of the envelope glycoproteins (Envs) of primate immunodeficiency viruses is, at least in part, regulated by endocytosis signal(s) located in the Env cytoplasmic domain. Here, we show that a membrane proximal signal that directs the simian immunodeficiency virus (SIV) Env to clathrin-coated pits, and is conserved in all SIV and human immunodeficiency virus Envs, conforms to a YxxØ motif (where x can be any amino acid and Ø represents a large hydrophobic residue). This motif is similar to that described for a number of cellular membrane proteins. By surface plasmon resonance we detected a high affinity interaction between peptides containing this membrane proximal signal and both AP1 and AP2 clathrin adaptor complexes. Mutation of the tyrosine in this membrane proximal motif in a SIV Env with a prematurely truncated cytoplasmic domain leads to a ] 25-fold increase in Env expression on infected cells. By contrast, the same mutation in an Env with a full-length cytoplasmic domain increases cell surface expression only 4-fold. We show that this effect results from the presence of additional endocytosis signals in the full-length cytoplasmic domain. Chimeras containing CD4 ecto-and membrane spanning domains and a full-length SIV Env cytoplasmic domain showed rapid endocytosis even when the membrane proximal tyrosine-based signal was disrupted. Mapping experiments indicated that at least some of the additional endocytosis information is located between residues 743 and 812 of Env from the SIV mac239 molecular clone. Together, our findings indicate that the cytoplasmic domain of SIV Env contains multiple endocytosis and/or trafficking signals that modulate its surface expression on infected cells, and suggest an important role for this function in pathogenesis.

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Section: Determination Of Kinetic Rate Constantsmentioning

confidence: 99%

The Simian Immunodeficiency Virus Envelope Glycoprotein Contains Multiple Signals that Regulate its Cell Surface Expression and Endocytosis

Bowers

Pelchen–Matthews

Höning

et al. 2000

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“…Data analysis was performed by nonlinear least-squares analysis as described by O'Shannessy et al (1993) using the LevenbergMarquardt algorithm as implemented in the Origin 4.1 software (Microcal Software, Northampton, Massachusetts). Association and dissociation kinetics were both observed to be biphasic for the majority of the SK species studied.…”

Section: Surface Plasmon Resonancementioning

confidence: 99%

“…Dissociation and association SPR curves were fitted using the equations reported by O'Shannessy et al (1993). Dissociation curves were first fitted in order to obtain values for the two dissociation rate constants kd,] and for each concentration of analyte (Plg).…”

Section: Surface Plasmon Resonancementioning

confidence: 99%

Analysis of the interactions between streptokinase domains and human plasminogen

et al. 1998

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The contrasting roles of streptokinase (SK) domains in binding human Glul-plasminogen (Plg) have been studied using a set of proteolytic fragments, each of which encompasses one or more of SK's three structural domains (A, B, C). Direct binding experiments have been performed using gel filtration chromatography and surface plasmon resonance. The latter technique has allowed estimation of association and dissociation rate constants for interactions between Plg and intact SK or SK fragments. Each of the SK fragments that contains domain B (fragments A2-B-C, A2-B, B-C, and B) binds Plg with similar affinity, at a level approximately 100-to 1,000-fold lower than intact SK. Experiments using 10 mM 6-aminohexanoic acid or 50 mM benzamidine demonstrate that either of these two lysine analogues abolishes interaction of domain B with Plg. Isolated domain C does not show detectable binding to Plg. Moreover, the additional presence of domain C within other SK fragments (B-C and A2-B-C) does not alter significantly their affinities for Plg. In addition, Plg-binding by a noncovalent complex of two SK fragments that contains domains A and B is similar to that of domain B. By contrast, species containing domain B and both domains A and C (intact SK and the two-chain complex AI . A2-B-C) show a significantly higher affinity for Plg, which could not be completely inhibited by saturating amounts of 6-AHA. These results show that SK domain B interacts with Plg in a lysine-dependent manner and that although domains A and C do not appear independently to possess affinity for Plg, they function cooperatively to establish the additional interactions with Plg to form an efficient native-like Plg activator complex.Keywords: fibrinolysis; lysine-binding site; plasminogen; serine protease; streptokinase; zymogen activation Streptococcus equisimilis streptokinase (SK) is a single-chain secreted protein of 414 amino acids (Malke et al., 1985), which indirectly causes the activation of the blood plasma zymogen human plasminogen (Plg) to the fibrinolytic enzyme plasmin (Christensen, 1945) via active-site formation within Plg. This proceeds without proteolytic cleavage by means of a rapid and avid binding

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“…The biotinylated GutR binding site-containing fragment was immobilized in the second flow cell to 71 resonance units. With this level of the GutR binding site immobilized, no mass transport effect that would interfere with the kinetic measurement was observed (10). GutR in a binding buffer (10 mM Tris-HCl, pH 8.0, 150 mM NaCl, 0.05% surfactant P20, and 20 mM MgCl 2 ) was injected into the sensor chip and passed from the first flow cell to the second flow cell.…”

Section: Determination Of the Association And Dissociation Rate Constmentioning

confidence: 99%

“…In the first step, the binding rate, expressed in terms of dR/dt (where R represents the resonance unit), was plotted against the response (R). Based on the kinetic equation (10), such a plot should yield a straight line with the slope (designated as k s ) representing the value of…”

Section: Determination Of the Association And Dissociation Rate Constmentioning

confidence: 99%

Roles of Glucitol in the GutR-mediated Transcription Activation Process in Bacillus subtilis

Poon¹,

Chen

Wong

2001

Journal of Biological Chemistry

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Glucitol induction in Bacillus subtilis requires a transcription activator, GutR, and a sequence located upstream of the gut promoter. To understand the initial steps involved in the GutR-mediated transcription activation process and the physiological roles of glucitol, GutR was overproduced and purified. In the absence of glucitol, GutR exists as a monomer and binds directly to its binding site in the gut regulatory region. This binding site was mapped to a 29-base pair imperfect inverted repeat located between ؊78 and ؊50, and there is only one GutR binding site within the regulatory region. The kinetic parameters of the interaction between GutR and its binding site were monitored in real time using surface plasmon resonance. The half-life of the GutR-DNA complex in the absence of glucitol was estimated to be 6.8 min. In contrast, in the presence of glucitol, the halflife of the complex was extended to longer than 19 h by affecting only the off-rate but not the on-rate. This effect is glucitol-specific. These data indicate that glucitol binds to GutR and induces GutR to have an extremely tight binding at its binding site. The physiological relevance of this process in transcription activation is discussed.With the addition of glucitol to Bacillus subtilis, a set of glucitol-inducible genes is selectively turned on (1). These inducible genes include gutA-and gutB-encoding glucitol permease and glucitol dehydrogenase, respectively. They are the members of the gut (glucitol utilization) operon and are arranged in the order from gutB to gutA. This operon is subject to both negative and positive regulations at the transcription level with the positive regulation playing a major role in the induction process. The regulatory region of the gut operon can be divided into three parts (2). The central portion is an unusual promoter with its Ϫ10 and Ϫ35 elements similar to those recognized by the major RNA polymerase (E A ) separated from each other by a 15-bp 1 spacer rather than a typical 17-bpspacer. An inverted repeat sequence located downstream of the promoter serves as a negative regulatory element (2). Although deletion of this sequence does not result in the constitutive expression of the gut operon in the absence of glucitol, the induced expression from this system increases by almost 4-fold. A region (from Ϫ126 to Ϫ48 with the transcription start site as ϩ1) located upstream of the promoter is required for glucitol induction and is suggested to contain binding sites for a transcription activator, GutR. The structural gene encoding GutR is located upstream of the gut operon and is transcribed in an opposite direction relative to that of the gut operon (3, 4

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[15] Determination of rate and equilibrium binding constants for macromolecular interactions by surface plasmon resonance

Cited by 130 publications

References 17 publications

The Simian Immunodeficiency Virus Envelope Glycoprotein Contains Multiple Signals that Regulate its Cell Surface Expression and Endocytosis

The Simian Immunodeficiency Virus Envelope Glycoprotein Contains Multiple Signals that Regulate its Cell Surface Expression and Endocytosis

Analysis of the interactions between streptokinase domains and human plasminogen

Roles of Glucitol in the GutR-mediated Transcription Activation Process in Bacillus subtilis

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