Expression and purification of human tryptophan hydroxylase from Escherichia coli and Pichia pastoris

McKinney, Jeffrey A.; Knappskog, Per M.; Pereira, Jacinto; Ekern, Trude; Toska, Karen; Kuitert, Baukje B; Levine, David; Gronenborn, Angela M.; Martı́nez, Aurora; Haavik, Jan

doi:10.1016/j.pep.2003.09.014

Cited by 38 publications

(67 citation statements)

References 36 publications

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“…Expression of Tph1 has also been reported in the raphe neurons during a late developmental phase, peaking at P21 (Nakamura et al, 2006), although this was not confirmed in a more recent study (Gutknecht et al, 2009). Studies of the kinetics of both enzymes have shown that Tph1 has a higher affinity for tryptophan and a higher enzymatic activity than Tph2 (McKinney et al, 2004;Nakamura et al, 2006).…”

Section: Targeting 5-ht Synthesis: Tryptophan Hydroxylase Knockout Micementioning

confidence: 92%

Genetic Models of Serotonin (5‐HT) Depletion: What do They Tell Us About the Developmental Role of 5‐HT?

Trowbridge¹,

Narboux-Nême²,

Gaspar³

2010

The Anatomical Record

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A large number of hyposerotonergic genetic models have been generated over the past few years. Serotonin (5-HT) depletion has been obtained via targeting of genes involved in 5-HT synthesis (Tph1 and Tph2), specification and determination of the 5-HT phenotype during development (GATA3, Pet1, and Lmx1b), and 5-HT storage or clearance (Vmat2 and SERT). Here we review these various models from a developmental perspective, beginning with a description of the sources of 5-HT during development. We then summarize the neurological and behavioral alterations that have been observed in the genetic hyposerotonergic models. Although these models appear to have normal brain development and do not exhibit any gross morphological defects, problems in somatic growth and physiological functions have been observed. Abnormal adult behavior is also seen, although whether it results from depletion of 5-HT during development or functional 5-HT deficiencies in adult life remains unclear. Evidence from these hyposerotonergic models suggests that the developing brain may not need 5-HT for the establishment of general organization and structure. However, central 5-HT appears to be necessary for postnatal body growth, maturation of respiratory and vegetative control, and possibly for the development of normal adult behavior. Anat Rec, 294:1615Rec, 294: -1623Rec, 294: , 2011. V V C 2011 Wiley-Liss, Inc.

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Section: Targeting 5-ht Synthesis: Tryptophan Hydroxylase Knockout Micementioning

confidence: 92%

Genetic Models of Serotonin (5‐HT) Depletion: What do They Tell Us About the Developmental Role of 5‐HT?

Trowbridge¹,

Narboux-Nême²,

Gaspar³

2010

The Anatomical Record

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“…The selectivity of the TPHs for pterins with methyl substituents at the N8 position is currently being investigated using purified recombinant human TPH1 and TPH2. In a comparative study of various pteridine analogs, we have recently observed that 8-methyl-6,7-dimethyl-5,6,7,8-tetrahydropterin (8-M-6,7-DMPH4) acts as an inhibitor of both TPH1 and TPH2 (up to 30% inhibition with 5 µM 8-M-6,7-DMPH4 in our standard TPH assay mixture) [121], while no inhibition of human PAH or TH1 activities was measured at the same concentration (un Much effort was put into the development of substrate analogs capable of specific inhibition of TH in the early 1960's. The motivation underlying the work of the groups Fig (5).…”

Section: The Iron Sitementioning

confidence: 95%

Selectivity and Affinity Determinants for Ligand Binding to the Aromatic Amino Acid Hydroxylases

Teigen

McKinney²,

Haavik³

et al. 2007

CMC

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Hydroxylation of the aromatic amino acids phenylalanine, tyrosine and tryptophan is carried out by a family of non-heme iron and tetrahydrobiopterin (BH4) dependent enzymes, i.e. the aromatic amino acid hydroxylases (AAHs). The reactions catalyzed by these enzymes are important for biomedicine and their mutant forms in humans are associated with phenylketonuria (phenylalanine hydroxylase), Parkinson's disease and DOPA-responsive dystonia (tyrosine hydroxylase), and possibly neuropsychiatric and gastrointestinal disorders (tryptophan hydroxylase 1 and 2). We attempt to rationalize current knowledge about substrate and inhibitor specificity based on the three-dimensional structures of the enzymes and their complexes with substrates, cofactors and inhibitors. In addition, further insights on the selectivity and affinity determinants for ligand binding in the AAHs were obtained from molecular interaction field (MIF) analysis. We applied this computational structural approach to a rational analysis of structural differences at the active sites of the enzymes, a strategy that can help in the design of novel selective ligands for each AAH.

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“…One of the advantages of this phenotype is that low growth rates may be desirable for production of certain recombinant products [26,27]. Currently, the majority of researchers use the Mut + phenotype [47,102,129], although some researchers are also using the Mut s phenotype [1,71,83,85,126].…”

Section: Methanol Utilization Phenotypesmentioning

confidence: 99%

Heterologous protein production using the Pichia pastoris expression system

Macauley-Patrick

Fazenda

McNeil

et al. 2005

Yeast

1,095

633

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The Pichia pastoris expression system is being used successfully for the production of various recombinant heterologous proteins. Recent developments with respect to the Pichia expression system have had an impact on not only the expression levels that can be achieved, but also the bioactivity of various heterologous proteins. We review here some of these recent developments, as well as strategies for reducing proteolytic degradation of the expressed recombinant protein at cultivation, cellular and protein levels. The problems associated with post-translational modifications performed on recombinant proteins by P. pastoris are discussed, including the effects on bioactivity and function of these proteins, and some engineering strategies for minimizing unwanted glycosylations. We pay particular attention to the importance of optimizing the physicochemical environment for efficient and maximal recombinant protein production in bioreactors and the role of process control in optimizing protein production is reviewed. Finally, future aspects of the use of the P. pastoris expression system are discussed with regard to the production of complex membrane proteins, such as G protein-coupled receptors, and the industrial and clinical importance of these proteins.

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Expression and purification of human tryptophan hydroxylase from Escherichia coli and Pichia pastoris

Cited by 38 publications

References 36 publications

Genetic Models of Serotonin (5‐HT) Depletion: What do They Tell Us About the Developmental Role of 5‐HT?

Genetic Models of Serotonin (5‐HT) Depletion: What do They Tell Us About the Developmental Role of 5‐HT?

Selectivity and Affinity Determinants for Ligand Binding to the Aromatic Amino Acid Hydroxylases

Heterologous protein production using the Pichia pastoris expression system

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