title: Cysteine-rich LIM domains of LIM-homeodomain and LIM-only proteins contain zinc but not iron.
abstract: The structure of LIM domains has major implications for transcription because proteins such as Is1-1 contain two LIM domains associated with a homeodomain, and RBTN1/Ttg-1 and RBTN2/Ttg-2 contain two LIM domains but no homeodomain. Conserved cysteine and histidine residues in the LIM domains suggest a metal-binding role. RBTN and Is1-1 LIM proteins have been made in Escherichia coli and insect cell expression systems and their metal content has been determined using atomic absorption spectroscopy and electron paramagnetic resonance spectroscopy. LIM proteins expressed in soluble form contain zinc atoms, whereas bacterial inclusion bodies invariably also have Fe-S clusters. The latter are identified as linear [Fe3S4]+ clusters and appear to result from incorrect metal coordination by E. coli. These studies show that RBTN1, RBTN2, and Is1-1 are metalloproteins that contain zinc but not iron and, therefore, that the LIM domain represents a zinc-binding domain.
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Cambridge, United Kingdom.
authors.forename: V E
authors.initials: VE
authors.lastname: Archer
authors.forename: J
authors.initials: J
authors.lastname: Breton
authors.forename: I
authors.initials: I
authors.lastname: Sanchez-Garcia
authors.forename: H
authors.initials: H
authors.lastname: Osada
authors.forename: A
authors.initials: A
authors.lastname: Forster
authors.forename: A J
authors.initials: AJ
authors.lastname: Thomson
authors.forename: T H
authors.initials: TH
authors.lastname: Rabbitts
mesh_terms: D003545:Cysteine / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000737:chemistry*; D005801:Genes, Homeobox; D018398:Homeodomain Proteins; D007501:Iron / Q000737:chemistry; D008667:Metalloproteins / Q000737:chemistry*; D015513:Oncogene Proteins; D011993:Recombinant Fusion Proteins; D012995:Solubility; D015032:Zinc / Q000737:chemistry; D016335:Zinc Fingers
chemical_list: D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; C085115:Is1-1 protein; D008667:Metalloproteins; D015513:Oncogene Proteins; D011993:Recombinant Fusion Proteins; D007501:Iron; D015032:Zinc; D003545:Cysteine