Expression of dopamine beta-hydroxylase in Drosophila Schneider 2 cells. Evidence for a mechanism of membrane binding other than uncleaved signal peptide

Gibson, Keisha; Vanek, Philip G.; Kaloss, William D.; Collier, Galen; Connaughton, John F.; Angelichio, Monica; Livi, George P.; Fleming, Patrick

doi:10.1016/s0021-9258(18)98377-8

Cited by 24 publications

(8 citation statements)

References 51 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…14, lanes 2 and 4). LI-cadherin expressed in $2 cells is, hence, N-glycosylated in accordance with previous reports that Drosophila cells can synthesize and transfer N-linked oligosaccharides onto vertebrate heterologous expressed glycoproteins (Domingo and Trowbridge, 1988;Gibson et al, 1993). The lower molecular mass of 109 kD of LI-cadherin expressed in $2 cells most likely reflects that insect cells are unable to elongate trimmed oligosaccharides to complex side chains (Butters et al, 1981;Hsieh and Robbins, 1984;Kuroda et al, 1990).…”

Section: Heterologous Expression Of Li-cadherin In Drosophila $2 Cellssupporting

confidence: 88%

Liver-intestine cadherin: molecular cloning and characterization of a novel Ca(2+)-dependent cell adhesion molecule expressed in liver and intestine.

Berndorff

Geßner

Kreft

et al. 1994

The Journal of Cell Biology

173

179

View full text Add to dashboard Cite

Abstract.A novel member of the cadherin family of cell adhesion molecules has been characterized by cloning from rat liver, sequencing of the corresponding cDNA, and functional analysis after heterologous expression in nonadhesive $2 cells, cDNA clones were isolated using a polyclonal antibody inhibiting Ca2+-dependent intercellular adhesion of hepatoma cells. As inferred from the deduced amino acid sequence, the novel molecule has homologies with E-, P-, and N-cadherins, but differs from these classical cadherins in four characteristics. Its extracellular domain is composed of five homologous repeated domains instead of four characteristic for the classical cadherins. Four of the five domains are characterized by the sequence motifs DXNDN and DXD or modifications thereof representing putative Ca2+-binding sites of classical cadherins. In its NH2-terminal region, this cadherin lacks both the precursor segment and the endogenous protease cleavage site RXKR found in classical cadherins. In the extracellular EC1 domain, the novel cadherin contains an AAL sequence in place of the HAV sequence motif representing the common cell adhesion recognition sequence of E-, P-, and N-cadherin. In contrast to the conserved cytoplasmic domain of classical cadherins with a length of 150-160 amino acid residues, that of the novel cadherin has only 18 amino acids. Examination of transfected $2 cells showed that despite these structural differences, this cadherin mediates intercellular adhesion in a Ca2+-dependent manner. The novel cadherin is solely expressed in liver and intestine and was, hence, assigned the name LI-cadherin. In these tissues, LI-cadherin is localized to the basolateral domain of hepatocytes and enterocytes. These results suggest that LI-cadherin represents a new cadherin subtype and may have a role in the morphological organization of liver and intestine.C ELL-cell interactions are of fundamental importance for the development and the maintenance of tissues and organs in multicellular organisms. The basic morphogenetic processes involved in organogenesis, including cellular aggregation, segregation, and migration, are mediated and controlled by an increasingly large and complex number of cell adhesion molecules that exhibit a well-regulated spatiotemporal pattern of expression during development and regeneration (for review see Simons and Fuller, 1985;Ekblom et al

show abstract

Section: Heterologous Expression Of Li-cadherin In Drosophila $2 Cellssupporting

confidence: 88%

Liver-intestine cadherin: molecular cloning and characterization of a novel Ca(2+)-dependent cell adhesion molecule expressed in liver and intestine.

Berndorff

Geßner

Kreft

et al. 1994

The Journal of Cell Biology

173

179

View full text Add to dashboard Cite

show abstract

“…DβH catalyzes the benzylic hydroxylation of dopamine ( 6 ) to norepinephrine ( 7 ), an essential small-molecule neurotransmitter modulating behavioral and physiological processes (Scheme A). Kobayashi and co-workers expressed human DβH in transgenic mice and Gibson and co-workers were able to express DβH in Drosophila melanogaster Schneider 2 (S2) cells . The insect homologue tyramine β-hydroxylase TBM was also expressed in Schneider 2 (S2) cells and a Bombyx mori strain. , Recombinant human DβH expressed in yeast or E.…”

Section: Copper-dependent Enzymesmentioning

confidence: 99%

“…Kobayashi and co-workers expressed human DβH in transgenic mice 96 and Gibson and co-workers were able to express DβH in Drosophila melanogaster Schneider 2 (S2) cells. 97 The insect homologue tyramine β-hydroxylase TBM was also expressed in Schneider 2 (S2) cells and a Bombyx mori strain. 62,98 Recombinant human DβH expressed in yeast or E. coli can be purchased from different biotechnological companies, such as FisherScientific, Cusabio, or LSBio.…”

Section: ■ Metal-free Enzymesmentioning

confidence: 99%

Enzymatic Hydroxylations of sp³-Carbons

et al. 2021

View full text Add to dashboard Cite

Enzymatic hydroxylation of activated and nonactivated sp 3 -carbons attracts keen interest from the chemistry community as it is one of the most challenging tasks in organic synthesis. Nature provides a vast number of enzymes with an enormous catalytic versatility to fulfill this task. Given that those very different enzymes have a distinct specificity in substrate scope, selectivity, activity, stability, and catalytic cycle, it is interesting to outline similarities and differences. In this Review, we intend to delineate which enzymes possess considerable advantages within specific issues. Heterologous production, crystal structure availability, enzyme engineering potential, and substrate promiscuity are essential factors for the applicability of these biocatalysts.

show abstract

“…The polypeptide sequence of the bovine enzyme has been determined by two groups (Robertson etal., 1990(Robertson etal., ,1991Wang et al, 1990), and the genes for the human (Lamouroux et al, 1987;Kobayashiet al, 1989),bovine (Taljanidiszetal., 1989;Wang et al, 1990;Lewis et al, 1990), rat (McMahon et al, 1990), and mouse (Nakano et al, 1992) enzyme have been cloned. The cloned bovine enzyme has been expressed in PC 12 cells (Lewis et al, 1992), COS cells (Ishii et al, 1991), and drosophila Schneider 2 cells (Gibson et al, 1993), but has not yet been overexpressed in large quantities. The successful expression of dopamine /3-hydroxylase holds promise for sitespecific mutagenesis and the prospect of even more detailed mechanistic studies.…”

mentioning

confidence: 99%

“…In the absence of any evidence for a transmembrane segment in the enzyme, Taljanidisz and co-workers proposed that the signal peptide may be the membrane anchor for membranebound dopamine /3-hydroxylase and provided N-terminal sequencing evidence that purified membrane-bound dopamine /3-hydroxylase contains the signal peptide (Taljanidisz et al, 1989). Investigations in another laboratory originally found no evidence of the signal peptide in membrane-bound enzyme , but later found that 20% of the purified membrane bound form contained signal peptide (Gibson et al, 1993). However, expression of dopamine /3-hydroxylase in insect cells has revealed that complete removal of the signal peptide does not result in total conversion to the soluble form (Gibson et al, 1993).…”

mentioning

confidence: 99%

Complete Assignment of Disulfide Bonds in Bovine Dopamine .beta.-Hydroxylase

Robertson

Adams²,

Medzihradszky³

et al. 1994

Biochemistry

View full text Add to dashboard Cite

Peptide mapping, chemical sequencing, microbore HPLC/electrospray ionization mass spectrometry (LC/ESI/MS), and matrix-assisted laser desorption mass spectrometry (MALDI/MS) were used to identify the sites of intra- and intermolecular disulfide linkages in bovine dopamine beta-hydroxylase. The enzyme contains 14 cysteines and seven disulfides per monomer. Edman sequencing of tryptic and peptic peptides determined linkages at positions Cys140-Cys582, Cys218-Cys269, Cys255-Cys281, Cys452-Cys474, Cys514-Cys514, and Cys516-Cys516, where cysteines at positions 514 and 516 on one monomer disulfide pair with their homologs on a second monomer. These linkages were confirmed by LC/ESI/MS and MALDI/MS. Further analysis by LC/ESI/MS and MALDI/MS identified linkages at positions Cys376-Cys489 and Cys380-Cys551. Cysteines 140 and 582 form a disulfide linkage that folds the C-terminus back in proximity to the N-terminus. The remaining intramolecular disulfides occur along two separate internal regions of the protein. The density of histidine residues in these two regions suggests binding sites for two Cu2+ atoms per monomer. In addition, previously identified amino acids that react with mechanism-based inactivators occur in these two regions. We propose that these five internal disulfide bonds define two Cu2+ binding domains that make up the active site of a dopamine beta-hydroxylase monomer. Considering previous data on the location of glycosylation sites, mechanism-based inactivation sites, and the disulfide linkages presented here, the data suggest an overall topology were the N- and C-termini are in close proximity and are solvent exposed and where the Cu2+ binding sites are buried in two interior domains stabilized by five disulfide bonds.

show abstract

Expression of dopamine beta-hydroxylase in Drosophila Schneider 2 cells. Evidence for a mechanism of membrane binding other than uncleaved signal peptide

Cited by 24 publications

References 51 publications

Liver-intestine cadherin: molecular cloning and characterization of a novel Ca(2+)-dependent cell adhesion molecule expressed in liver and intestine.

Liver-intestine cadherin: molecular cloning and characterization of a novel Ca(2+)-dependent cell adhesion molecule expressed in liver and intestine.

Enzymatic Hydroxylations of sp³-Carbons

Complete Assignment of Disulfide Bonds in Bovine Dopamine .beta.-Hydroxylase

Contact Info

Product

Resources

About

Expression of dopamine beta-hydroxylase in Drosophila Schneider 2 cells. Evidence for a mechanism of membrane binding other than uncleaved signal peptide

Cited by 24 publications

References 51 publications

Liver-intestine cadherin: molecular cloning and characterization of a novel Ca(2+)-dependent cell adhesion molecule expressed in liver and intestine.

Liver-intestine cadherin: molecular cloning and characterization of a novel Ca(2+)-dependent cell adhesion molecule expressed in liver and intestine.

Enzymatic Hydroxylations of sp3-Carbons

Complete Assignment of Disulfide Bonds in Bovine Dopamine .beta.-Hydroxylase

Contact Info

Product

Resources

About

Enzymatic Hydroxylations of sp³-Carbons