A determinant of substrate specificity predicted from the Acyl-Acyl carrier protein desaturase of developing cat's claw seed

Edgar B. Cahoon, Salehuzzaman Shah, John Shanklin, John Browse

Research output: Contribution to journalArticle

73 Scopus citations

Abstract

Cat's claw (Doxantha unguis-cati L.) vine accumulates nearly 80% palmitoleic acid (16:1Δ9) plus cis-vaccenic acid (18:1Δ11) in its seed oil. To characterize the biosynthetic origin of these unusual fatty acids, cDNAs for acyl-acyl carrier protein (acyl-ACP) desaturases were isolated from developing cat's claw seeds. The predominant acyl-ACP desaturase cDNA identified encoded a polypeptide that is closely related to the stearoyl (Δ9-18:0)-ACP desaturase from castor (Ricinis commuais L.) and other species. Upon expression in Eschcrichia coli, the cat's claw polypeptide functioned as a Δ9 acyl-ACP desaturase but displayed a distinct substrate specificity for palmitate (16:0)-ACP rather than stearate (18:0)-ACP. Comparison of the predicted amino acid sequence of the cat's claw enzyme with that of the castor Δ9-18:0-ACP desaturase suggested that a single amino acid substitution (L118W) might account in large part for the differences in substrate specificity between the two desaturases. Consistent with this prediction, conversion of leucine-118 to tryptophan in the mature castor Δ9-18:0-ACP desaturase resulted in an 80-fold increase in the relative specificity of this enzyme for 16:0-ACP. The alteration in substrate specificity observed in the L118W mutant is in agreement with a crystallographic model of the proposed substrate-binding pocket of the castor Δ9-18:0-ACP desaturase.

Original languageEnglish (US)
Pages (from-to)593-598
Number of pages6
JournalPlant physiology
Volume117
Issue number2
DOIs
StatePublished - Jan 1 1998

ASJC Scopus subject areas

  • Physiology
  • Genetics
  • Plant Science

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