Purification and partial sequence analysis of human T-cell growth factor.

Robb, Richard J.; Kutny, Rusty; Chowdhry, Vinay

doi:10.1073/pnas.80.19.5990

Cited by 117 publications

(59 citation statements)

References 24 publications

(23 reference statements)

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“…Samples for amino acid analysis were hydrolyzed in 6 M HCl at 1 lOC for 24, 48, and 72 h. To quantitate cysteine, samples were reduced with DTT, alkylated with iodoacetic acid, and rechromatographed (26). Amino acid analysis was performed on a Beckman 6300 ion exchange/ ninhydrin analyzer.…”

Section: Amino Acid Analysis and N-terminal Sequencingmentioning

confidence: 99%

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Cytochrome P-450 from the Mesocarp of Avocado (Persea americana)

O’Keefe

Leto²

1989

Plant Physiol.

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The microsomal fraction from the mesocarp of avocado (Persea amerkana) is one of few identified rich sources of plant cytochrome P-450. Cytochrome P-450 from this tissue has been solubilized and purified. Enzymatic assays (p-chloro-N-methylaniline demethylase) and spectroscopic observations of substrate binding suggest a low spin form of the cytochrome, resembling that in the microsomal membrane, can be recovered. However, this preparation of native protein is a mixture of nearly equal proportions of two cytochrome P-450 polypeptides that have been resolved only under denaturing conditions. Overall similarities between these polypeptides include indistinguishable amino acid compositions, similar trypsin digest pattems, and cross reactivity with the same antibody. The amino terminal sequences of both polypeptides are identical, with the exception that one of them lacks a methionine residue at the amino terminus. This sequence exhibits some similarites with the membrane targeting signal found at the amino terminus of most mammalian cytochromes P-450.Cytochrome P450 dependent monooxygenases are widespread in nature. They are involved in a variety of catabolic and biosynthetic pathways and in the metabolism of drugs and xenobiotics. In plants, Cyt P-450 has been identified, for example, in the trans-cinnamic acid and kaurene hydroxylases (10,11). Others are suspected participants in a number of reactions associated with xenobiotic metabolism (6, 8) (for reviews see Refs. 4 and 20). Within these monooxygenase systems, Cyt P-450 serves as a terminal oxidase, responsible for substrate recognition, binding, and oxygen redox chemistry (5, 29).The mechanistic and structural details of Cyt P450 dependent monooxygenases have been developed largely from studies of the bacterial (Pseudomonas putida) camphor hydroxylase system (23,29). Because of their importance in xenobiotic and drug metabolism, these enzymes have also been thoroughly studied in mammalian liver (3,5,31). In contrast, Cyt P-450 of higher plant origin has not been the subject of extensive biochemical characterization, primarily because of the low content in many plant tissues, difficulties with identification in Chl containing tissues, and supposed lability in homogenized plant extracts. ' (6,10,12,13,19,20). Cyt P-450 has been purified from tulip (Tulipa gesneriana) bulbs (12), and from tubers of Jerusalem artichoke (Helianthus tuberosus) (10). Additionally, an NADPH:Cyt P450 reductase has been purified from Jerusalem artichoke (2), and the related Cyt b5 and NADH:Cyt b5 reductase have been purified from Pisum sativum (16). Unfortunately, the tulip bulb Cyt P450 has not been associated with any enzymic activity, and polyclonal antibodies raised against it were not cross reactive with proteins from other species (1 1). The Jerusalem artichoke preparation can be reconstituted into its trans-cinnamate hydroxylase activity, but it has a low heme specific content and is not homogeneous (10). No primary sequence information has been reported for either o...

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Section: Amino Acid Analysis and N-terminal Sequencingmentioning

confidence: 99%

“…Amino acid analysis was performed on a Beckman 6300 ion exchange/ ninhydrin analyzer. N-terminal sequencing was done by automated Edman degradation (26).…”

Section: Amino Acid Analysis and N-terminal Sequencingmentioning

confidence: 99%

Cytochrome P-450 from the Mesocarp of Avocado (Persea americana)

O’Keefe

Leto²

1989

Plant Physiol.

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“…Early studies on human IL-2 protein were restricted to the protein purified from human T-cell leukemia cell line (Jurkat) (Robb et al, 1983;Stern et al, 1984). After recombinant protein technology, IL-2 was expressed after isolation of IL-2 mRNA from human leukemic T-cell line, followed by cloning of the complementary cDNA into a suitable vector.…”

Section: Discussionmentioning

confidence: 99%

Expression and purification of human IL-2 protein from Escherichia coli

Hassan¹,

El‐Mowafy²,

Zaher³

2018

Afr. J. Biotechnol.

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“…Amino acid sequencing of the Jurkat-derived material confirmed the primary structure of the protein, deduced from the cDNA sequence, and established the N-terminal residues after signal peptide cleavage (12,13). IL-2 appears to undergo some posttranslational modifications that account for the heterogeneity observed in size and charge (14).…”

mentioning

confidence: 95%

“…Natural IL-2 has been purified to homogeneity from the human T-cell leukemia line Jurkat (11,12). Amino acid sequencing of the Jurkat-derived material confirmed the primary structure of the protein, deduced from the cDNA sequence, and established the N-terminal residues after signal peptide cleavage (12,13).…”

mentioning

confidence: 99%

Modification and secretion of human interleukin 2 produced in insect cells by a baculovirus expression vector.

Smith¹,

Ju²,

Ericson³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

241

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A cDNA coding for human interleukin 2 (IL-2) was inserted into the genome of Autographa californica nuclear polyhedrosis virus adjacent to the polyhedrin promoter. Cells infected with recombinant virus produced high levels of Mr 15,500 IL-2 polypeptide, the majority of which was secreted into the culture medium during infection. The recombinant IL-2 was able to stimulate the growth of an IL-2-dependent cell line. The N-terminal amino acid sequence of the insect-derived IL-2 was identical to that of natural IL-2. Thus, a mammalian signal peptide was recognized and properly removed in insect cells.

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Purification and partial sequence analysis of human T-cell growth factor.

Cited by 117 publications

References 24 publications

Cytochrome P-450 from the Mesocarp of Avocado (Persea americana)

Cytochrome P-450 from the Mesocarp of Avocado (Persea americana)

Expression and purification of human IL-2 protein from Escherichia coli

Modification and secretion of human interleukin 2 produced in insect cells by a baculovirus expression vector.

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