A novel 4-methylideneimidazole-5-one-containing tyrosine aminomutase in enediyne antitumor antibiotic C-1027 biosynthesis

Steven D. Christenson, Wen Liu, Michael D. Toney, Ben Shen

Research output: Contribution to journalArticle

98 Citations (Scopus)

Abstract

The C-1027 enediyne antibiotic contains an unusual 3-chloro-4,5-dihydroxy-β-phenylalanine moiety that is thought to be derived from tyrosine by an aminomutase reaction. However, none of the genes identified within the C-1027 gene cluster encode proteins with strong homology to known aminomutases. The sgcC4 gene encodes a protein with strong homology to dehydroalanine-dependent histidine/phenylalanine ammonia lyases. The sgcC4 gene was expressed in E. coli, and overproduced SgcC4 was purified as a His6-tagged fusion protein. Biochemical characterization of the purified SgcC4 establishes that SgcC4 is an aminomutase that catalyzes the conversion of l-tyrosine to (S)-β-tyrosine and employs 4-methylideneimidazole-5-one (MIO) at its active site. The latter was supported by borohydride and cyanide inhibition studies and confirmed by site-directed mutagenesis. The S153A mutant exhibited a 340-fold decrease in kcat/KM. SgcC4 represents a novel type of aminomutase, extending the known MIO chemistry from ammonia lyases into aminomutases.

Original languageEnglish (US)
Pages (from-to)6062-6063
Number of pages2
JournalJournal of the American Chemical Society
Volume125
Issue number20
DOIs
StatePublished - May 21 2003

Fingerprint

Enediynes
Biosynthesis
Antibiotics
Tyrosine
Genes
Anti-Bacterial Agents
Ammonia-Lyases
Histidine Ammonia-Lyase
Proteins
Phenylalanine Ammonia-Lyase
Ammonia
Borohydrides
Cyanides
Multigene Family
Site-Directed Mutagenesis
Phenylalanine
Mutagenesis
Catalytic Domain
Histidine
Escherichia coli

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

A novel 4-methylideneimidazole-5-one-containing tyrosine aminomutase in enediyne antitumor antibiotic C-1027 biosynthesis. / Christenson, Steven D.; Liu, Wen; Toney, Michael D.; Shen, Ben.

In: Journal of the American Chemical Society, Vol. 125, No. 20, 21.05.2003, p. 6062-6063.

Research output: Contribution to journalArticle

Christenson, Steven D. ; Liu, Wen ; Toney, Michael D. ; Shen, Ben. / A novel 4-methylideneimidazole-5-one-containing tyrosine aminomutase in enediyne antitumor antibiotic C-1027 biosynthesis. In: Journal of the American Chemical Society. 2003 ; Vol. 125, No. 20. pp. 6062-6063.
@article{73699b7a693448cc81fa13000bf59dec,
title = "A novel 4-methylideneimidazole-5-one-containing tyrosine aminomutase in enediyne antitumor antibiotic C-1027 biosynthesis",
abstract = "The C-1027 enediyne antibiotic contains an unusual 3-chloro-4,5-dihydroxy-β-phenylalanine moiety that is thought to be derived from tyrosine by an aminomutase reaction. However, none of the genes identified within the C-1027 gene cluster encode proteins with strong homology to known aminomutases. The sgcC4 gene encodes a protein with strong homology to dehydroalanine-dependent histidine/phenylalanine ammonia lyases. The sgcC4 gene was expressed in E. coli, and overproduced SgcC4 was purified as a His6-tagged fusion protein. Biochemical characterization of the purified SgcC4 establishes that SgcC4 is an aminomutase that catalyzes the conversion of l-tyrosine to (S)-β-tyrosine and employs 4-methylideneimidazole-5-one (MIO) at its active site. The latter was supported by borohydride and cyanide inhibition studies and confirmed by site-directed mutagenesis. The S153A mutant exhibited a 340-fold decrease in kcat/KM. SgcC4 represents a novel type of aminomutase, extending the known MIO chemistry from ammonia lyases into aminomutases.",
author = "Christenson, {Steven D.} and Wen Liu and Toney, {Michael D.} and Ben Shen",
year = "2003",
month = "5",
day = "21",
doi = "10.1021/ja034609m",
language = "English (US)",
volume = "125",
pages = "6062--6063",
journal = "Journal of the American Chemical Society",
issn = "0002-7863",
publisher = "American Chemical Society",
number = "20",

}

TY - JOUR

T1 - A novel 4-methylideneimidazole-5-one-containing tyrosine aminomutase in enediyne antitumor antibiotic C-1027 biosynthesis

AU - Christenson, Steven D.

AU - Liu, Wen

AU - Toney, Michael D.

AU - Shen, Ben

PY - 2003/5/21

Y1 - 2003/5/21

N2 - The C-1027 enediyne antibiotic contains an unusual 3-chloro-4,5-dihydroxy-β-phenylalanine moiety that is thought to be derived from tyrosine by an aminomutase reaction. However, none of the genes identified within the C-1027 gene cluster encode proteins with strong homology to known aminomutases. The sgcC4 gene encodes a protein with strong homology to dehydroalanine-dependent histidine/phenylalanine ammonia lyases. The sgcC4 gene was expressed in E. coli, and overproduced SgcC4 was purified as a His6-tagged fusion protein. Biochemical characterization of the purified SgcC4 establishes that SgcC4 is an aminomutase that catalyzes the conversion of l-tyrosine to (S)-β-tyrosine and employs 4-methylideneimidazole-5-one (MIO) at its active site. The latter was supported by borohydride and cyanide inhibition studies and confirmed by site-directed mutagenesis. The S153A mutant exhibited a 340-fold decrease in kcat/KM. SgcC4 represents a novel type of aminomutase, extending the known MIO chemistry from ammonia lyases into aminomutases.

AB - The C-1027 enediyne antibiotic contains an unusual 3-chloro-4,5-dihydroxy-β-phenylalanine moiety that is thought to be derived from tyrosine by an aminomutase reaction. However, none of the genes identified within the C-1027 gene cluster encode proteins with strong homology to known aminomutases. The sgcC4 gene encodes a protein with strong homology to dehydroalanine-dependent histidine/phenylalanine ammonia lyases. The sgcC4 gene was expressed in E. coli, and overproduced SgcC4 was purified as a His6-tagged fusion protein. Biochemical characterization of the purified SgcC4 establishes that SgcC4 is an aminomutase that catalyzes the conversion of l-tyrosine to (S)-β-tyrosine and employs 4-methylideneimidazole-5-one (MIO) at its active site. The latter was supported by borohydride and cyanide inhibition studies and confirmed by site-directed mutagenesis. The S153A mutant exhibited a 340-fold decrease in kcat/KM. SgcC4 represents a novel type of aminomutase, extending the known MIO chemistry from ammonia lyases into aminomutases.

UR - http://www.scopus.com/inward/record.url?scp=0038288848&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0038288848&partnerID=8YFLogxK

U2 - 10.1021/ja034609m

DO - 10.1021/ja034609m

M3 - Article

C2 - 12785829

AN - SCOPUS:0038288848

VL - 125

SP - 6062

EP - 6063

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

SN - 0002-7863

IS - 20

ER -