Inactivation of Staphylococcal Nuclease by the Binding of Antibodies to a Distinct Antigenic Determinant

David H. Sachs; Alan N. Schechter; Ann Eastlake; Christian B. Anfinsen

doi:10.1021/bi00773a012

Inactivation of Staphylococcal Nuclease by the Binding of Antibodies to a Distinct Antigenic Determinant

David H. Sachs, Alan N. Schechter, Ann Eastlake, Christian B. Anfinsen

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Abstract

The interaction of antibodies with a distinct antigenic determinant in the region (99-126) of staphylococcal nuclease has been found to produce a soluble, inactive antibody-nuclease complex. The reaction has been followed spectrophotometrically, using the rate of hydrolysis of substrate DNA as a measure of residual, free nuclease concentration. This analysis has provided the following kinetic and equilibrium constants for the antibody-antigen interaction: Kass = 8.3 X 108 M-1, Kon =4.1 X 105 m-1 sec-1, A-offf t = 4.9 X 10-4 sec-1. Conversely, measurement of changes in nuclease activity can be used as a rapid and sensitive assay of antibody concentration. The Scatchard plot of the equilibrium inactivation data bends sharply near r = 1, which may reflect heterogeneity of the antibodies or may indicate possible steric interference in the binding of a second nuclease molecule to the bivalent antibody molecule.

Original language	English
Pages (from-to)	4268-4273
Number of pages	6
Journal	Biochemistry
Volume	11
Issue number	23
DOIs	https://doi.org/10.1021/bi00773a012
State	Published - 1 Nov 1972

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title = "Inactivation of Staphylococcal Nuclease by the Binding of Antibodies to a Distinct Antigenic Determinant",

abstract = "The interaction of antibodies with a distinct antigenic determinant in the region (99-126) of staphylococcal nuclease has been found to produce a soluble, inactive antibody-nuclease complex. The reaction has been followed spectrophotometrically, using the rate of hydrolysis of substrate DNA as a measure of residual, free nuclease concentration. This analysis has provided the following kinetic and equilibrium constants for the antibody-antigen interaction: Kass = 8.3 X 108 M-1, Kon =4.1 X 105 m-1 sec-1, A-offf t = 4.9 X 10-4 sec-1. Conversely, measurement of changes in nuclease activity can be used as a rapid and sensitive assay of antibody concentration. The Scatchard plot of the equilibrium inactivation data bends sharply near r = 1, which may reflect heterogeneity of the antibodies or may indicate possible steric interference in the binding of a second nuclease molecule to the bivalent antibody molecule.",

author = "Sachs, {David H.} and Schechter, {Alan N.} and Ann Eastlake and Anfinsen, {Christian B.}",

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doi = "10.1021/bi00773a012",

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journal = "Biochemistry",

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T1 - Inactivation of Staphylococcal Nuclease by the Binding of Antibodies to a Distinct Antigenic Determinant

AU - Sachs, David H.

AU - Schechter, Alan N.

AU - Eastlake, Ann

AU - Anfinsen, Christian B.

PY - 1972/11/1

Y1 - 1972/11/1

N2 - The interaction of antibodies with a distinct antigenic determinant in the region (99-126) of staphylococcal nuclease has been found to produce a soluble, inactive antibody-nuclease complex. The reaction has been followed spectrophotometrically, using the rate of hydrolysis of substrate DNA as a measure of residual, free nuclease concentration. This analysis has provided the following kinetic and equilibrium constants for the antibody-antigen interaction: Kass = 8.3 X 108 M-1, Kon =4.1 X 105 m-1 sec-1, A-offf t = 4.9 X 10-4 sec-1. Conversely, measurement of changes in nuclease activity can be used as a rapid and sensitive assay of antibody concentration. The Scatchard plot of the equilibrium inactivation data bends sharply near r = 1, which may reflect heterogeneity of the antibodies or may indicate possible steric interference in the binding of a second nuclease molecule to the bivalent antibody molecule.

AB - The interaction of antibodies with a distinct antigenic determinant in the region (99-126) of staphylococcal nuclease has been found to produce a soluble, inactive antibody-nuclease complex. The reaction has been followed spectrophotometrically, using the rate of hydrolysis of substrate DNA as a measure of residual, free nuclease concentration. This analysis has provided the following kinetic and equilibrium constants for the antibody-antigen interaction: Kass = 8.3 X 108 M-1, Kon =4.1 X 105 m-1 sec-1, A-offf t = 4.9 X 10-4 sec-1. Conversely, measurement of changes in nuclease activity can be used as a rapid and sensitive assay of antibody concentration. The Scatchard plot of the equilibrium inactivation data bends sharply near r = 1, which may reflect heterogeneity of the antibodies or may indicate possible steric interference in the binding of a second nuclease molecule to the bivalent antibody molecule.

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JO - Biochemistry

JF - Biochemistry

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Inactivation of Staphylococcal Nuclease by the Binding of Antibodies to a Distinct Antigenic Determinant

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