A procedure for amino acid sequencing in internal regions of proteins

Itai Benhar; Hanna Engelberg-Kulka

doi:10.1016/0378-1119(91)90394-Q

A procedure for amino acid sequencing in internal regions of proteins

Itai Benhar, Hanna Engelberg-Kulka^*

^*Corresponding author for this work

Hebrew University of Jerusalem

Research output: Contribution to journal › Article › peer-review

7 Scopus citations

Abstract

We describe a novel procedure for determining the amino acid (aa) sequence of the internal regions of proteins. This procedure has been implemented by directly determining the sequence of aa 65-75 of the product of the trpR gene of Escherichia coli, the trp repressor. This method is based on the insertion of the cleavage site of a specific protease (factor Xa) into the protein immediately before the region to be sequenced by Edman degradation. The simplicity of the procedure makes it appealing for studies of protein structure-function relationships, and of the expression of genetic information. The method is particularly useful when there is ambiguity concerning the co-linearity of the aa and nucleotide sequences.

Original language	English
Pages (from-to)	79-82
Number of pages	4
Journal	Gene
Volume	103
Issue number	1
DOIs	https://doi.org/10.1016/0378-1119(91)90394-Q
State	Published - 15 Jul 1991
Externally published	Yes

Keywords

Amino acid sequence
Edman degradation
in vitro mutagenesis
specific proteolytic cleavage
trp repressor

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10.1016/0378-1119(91)90394-Q

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abstract = "We describe a novel procedure for determining the amino acid (aa) sequence of the internal regions of proteins. This procedure has been implemented by directly determining the sequence of aa 65-75 of the product of the trpR gene of Escherichia coli, the trp repressor. This method is based on the insertion of the cleavage site of a specific protease (factor Xa) into the protein immediately before the region to be sequenced by Edman degradation. The simplicity of the procedure makes it appealing for studies of protein structure-function relationships, and of the expression of genetic information. The method is particularly useful when there is ambiguity concerning the co-linearity of the aa and nucleotide sequences.",

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N2 - We describe a novel procedure for determining the amino acid (aa) sequence of the internal regions of proteins. This procedure has been implemented by directly determining the sequence of aa 65-75 of the product of the trpR gene of Escherichia coli, the trp repressor. This method is based on the insertion of the cleavage site of a specific protease (factor Xa) into the protein immediately before the region to be sequenced by Edman degradation. The simplicity of the procedure makes it appealing for studies of protein structure-function relationships, and of the expression of genetic information. The method is particularly useful when there is ambiguity concerning the co-linearity of the aa and nucleotide sequences.

AB - We describe a novel procedure for determining the amino acid (aa) sequence of the internal regions of proteins. This procedure has been implemented by directly determining the sequence of aa 65-75 of the product of the trpR gene of Escherichia coli, the trp repressor. This method is based on the insertion of the cleavage site of a specific protease (factor Xa) into the protein immediately before the region to be sequenced by Edman degradation. The simplicity of the procedure makes it appealing for studies of protein structure-function relationships, and of the expression of genetic information. The method is particularly useful when there is ambiguity concerning the co-linearity of the aa and nucleotide sequences.

KW - Amino acid sequence

KW - Edman degradation

KW - in vitro mutagenesis

KW - specific proteolytic cleavage

KW - trp repressor

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JO - Gene

JF - Gene

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A procedure for amino acid sequencing in internal regions of proteins

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Keywords

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