A novel procedure for the preparation of biologically active recombinant peptides using a cyanylation reaction

Koyama, Nobuyuki; Kuriyama, Masato; Amano, Nobuyuki; Nishimura, Osamu

doi:10.1016/0168-1656(94)90213-5

Cited by 10 publications

(11 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…PrRPs were chemically synthesized with an automatic peptide synthesizer (model 430, ABI). They were also prepared as recombinant peptides produced in Escherichia coli 25,26 . Synthetic bovine PrRP31 was labelled with 125 I by use of [ 125 I] Bolton-Hunter reagent (NEN/Dupont).…”

mentioning

confidence: 99%

Elastin is an essential determinant of arterial morphogenesis

Brooke²,

Davis

et al. 1998

Nature

711

546

View full text Add to dashboard Cite

Elastin, the main component of the extracellular matrix of arteries, was thought to have a purely structural role. Disruption of elastin was believed to lead to dissection of arteries, but we showed that mutations in one allele encoding elastin cause a human disease in which arteries are blocked, namely, supravalvular aortic stenosis. Here we define the role of elastin in arterial development and disease by generating mice that lack elastin. These mice die of an obstructive arterial disease, which results from subendothelial cell proliferation and reorganization of smooth muscle. These cellular changes are similar to those seen in atherosclerosis. However, lack of elastin is not associated with endothelial damage, thrombosis or inflammation, which occur in models of atherosclerosis. Haemodynamic stress is not associated with arterial obstruction in these mice either, as the disease still occurred in arteries that were isolated in organ culture and therefore not subject to haemodynamic stress. Disruption of elastin is enough to induce subendothelial proliferation of smooth muscle and may contribute to obstructive arterial disease. Thus, elastin has an unanticipated regulatory function during arterial development, controlling proliferation of smooth muscle and stabilizing arterial structure.

show abstract

mentioning

confidence: 99%

Elastin is an essential determinant of arterial morphogenesis

Brooke²,

Davis

et al. 1998

Nature

711

546

View full text Add to dashboard Cite

show abstract

“…All expression plasmids were constructed using pTF plasmid, a derivative of pTB960-7, as described previously. 14 The cDNA fragments of human apelin, PrRPs (bovine, rat and human) and GALPs (porcine, human and rat) were prepared by the annealing of synthetic oligonucleotides and insertion into the cloning site of pTF plasmid.…”

Section: Construction Of Expression Plasmidsmentioning

confidence: 99%

“…We previously reported two efficient methods for obtaining recombinant peptides and proteins: a cysteine site-specific cleavage reaction of the fusion protein (Scheme 1); 14 and removal of an additional N-terminal methionine residue after transamination (Scheme 2). 15, 16 In the first method peptides are expressed in the form of fusion proteins with basic fibroblast growth factor mutein (CS23) as a fusion partner, which is easily purified by heparin affinity chromatography.…”

Section: Introductionmentioning

confidence: 99%

“…17 The SH groups of the denatured fusion proteins are then converted to SCN groups with 1-cyano-4-(dimethylamino)pyridinium tetrafluoroborate (DMAP-CN), and subsequent exposure to alkaline conditions results in cleavage at the amino group of the modified cysteine residue at the junction site, to yield the desired peptides. 14 Interestingly enough, this method can yield the Cterminal α-amide by treatment of the cyanylated protein with ammonia, 18 while the C-terminal α-acid can be obtained by treatment with sodium or potassium hydroxide. Cyanogen bromide is generally used as a site-specific cleavage reagent of the fusion proteins.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

A novel system for the preparation of orphan receptor ligand peptides

Nishimura

Koyama

Itoh

et al. 2001

J. Chem. Soc., Perkin Trans. 1

Self Cite

View full text Add to dashboard Cite

“…All of the expression plasmids were constructed using pTF plasmid, a derivative of pTB960-7. 4 Rat GALP and porcine GALP were inserted into the amino-and carboxy-terminal of CS23, respectively. Methionine at position 76 in CS23 was converted into leucine using QuickChange TM Site-Directed Mutagenesis Kit (Stratagene).…”

Section: Experimental Construction Of Expression Plasmidsmentioning

confidence: 99%