Role of N-Glycosylation in Human Angiotensinogen

Gimenez-Roqueplo, Anne-Paule; Célérier, Jérôme; Lucarelli, G.; Corvol, Pierre; Jeunemaı̂tre, Xavier

doi:10.1074/jbc.273.33.21232

Cited by 38 publications

(37 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In vivo glycosylation has been shown to alter the stability, intracellular transport, secretion, distribution, and overall activity of a variety of proteins (Hilgenfeldt, 1988;Mann et al, 1996;Gimenez-Roqueplo et al, 1998;Barrios-De-Tomasi et al, 2002). We, along with others, have hypothesized that glycosylation of smaller peptides may have desirable effects on the pharmacokinetic and/or pharmacodynamic properties of the parent peptide (Albert et al, 1993;Polt et al, 1994;Negri et al, 1998;Susaki et al, 1999;Suzuki et al, 1999a,b;Bilsky et al, 2000).…”

Section: Discussionmentioning

confidence: 99%

Antinociceptive Structure-Activity Studies with Enkephalin-Based Opioid Glycopeptides

Elmagbari¹,

Egleton²,

Palian³

et al. 2004

J Pharmacol Exp Ther

View full text Add to dashboard Cite

Development of opioid peptides as therapeutic agents has historically been limited due to pharmacokinetic issues including stability and blood-brain barrier (BBB) permeability. Glycosylation of opioid peptides can increase peptide serum stability and BBB penetration. To further define the requirements for optimizing in vivo antinociceptive potency following intravenous administration, we synthesized a series of enkephalin-based glycopeptides using solid phase 9-fluorenylmethyloxy carbamate methods. The compounds differed in the sixth and subsequent amino acid residues (Ser or Thr) and in the attached carbohydrate moiety. In vitro binding and functional smooth muscle bioassays indicated that the addition of mono-or disaccharides did not significantly affect the opioid receptor affinity or agonist activity of the glycopeptides compared with their unglycosylated parent peptides. All of the glycopeptides tested produced potent antinociceptive effects in male ICR mice following intracerebroventricular injection in the 55°C tail-flick test. The calculated A 50 values for the Ser/Thr and monosaccharide combinations were all very similar with values ranging from 0.02 to 0.09 nmol. Selected compounds were administered to mice intravenously and tested for antinociception to indirectly assess serum stability and BBB penetration. All compounds tested produced full antinociceptive effects with calculated A 50 values ranging from 2.2 to 46.4 mol/kg with the disaccharides having potencies that equaled or exceeded that of morphine on a micromoles per kilogram basis. Substitution of a trisaccharide or bis-and tris-monosaccharides resulted in a decrease in antinociceptive potency. These results provide additional support for the utility of glycosylation to increase central nervous system bioavailability of small peptides and compliment our ongoing stability and blood-brain barrier penetration studies.

show abstract

Section: Discussionmentioning

confidence: 99%

Antinociceptive Structure-Activity Studies with Enkephalin-Based Opioid Glycopeptides

Elmagbari¹,

Egleton²,

Palian³

et al. 2004

J Pharmacol Exp Ther

View full text Add to dashboard Cite

show abstract

“…Although the P3 0 residue in human angiotensinogen is also His, whereas it is Tyr in pig and rat angiotensinogens, the role of His13 of human angiotensinogen is probably different from that of His13 of sheep angiotensinogen due to the N-glycosylation of adjacent Asn14. 25) We have reported that Tyr83 of human renin also contributes to biphasic pH dependence and that it is essential to the catalytic reaction at basic pH. 10) Tyr83 constitutes the S3 0 subsite pocket.…”

Section: )mentioning

confidence: 99%

Ser84 of Human Renin Contributes to the Biphasic pH Dependence of the Renin-Angiotensinogen Reaction

Iwata

Nakagawa

Nishiuchi

et al. 2007

Bioscience, Biotechnology, and Biochemistry

View full text Add to dashboard Cite

“…Iodo (24) was analyzed by SDS-PAGE in non-reducing and reducing conditions (ϩdithiothreitol (DTT) 10 mM) after denaturation (4 min at 94°C). The gel was immediately stained using the colloidal blue stain kit (Novex).…”

Section: Methodsmentioning

confidence: 99%

“…The cells were transferred to serum-free medium (Dulbecco's modified Eagles's medium) 2 days after transfection and incubated for 48 h. The resulting supernatants containing recombinant proteins were analyzed on 9% SDS-PAGE with or without 2.5% ␤-mercaptoethanol and Western blotting (24). Angiotensinogen recombinants were detected with the anti-human angiotensinogen polyclonal antibody:HCL (diluted 1:6000) (28).…”

mentioning

confidence: 99%

“…It has been subcloned into the eucaryotic expression vector pECE (23). Site-directed mutagenesis of Met 235 to Thr and of each Cys (at positions 18, 138, 232, and 308) to Ala was performed with the same procedure previously described (24). One of the following antisense mutagenesis oligonucleotide was used: (Met 235 -Thr) 5Ј-GTCCACACTGGCTCCCGTCAGGGAGCAGCCAGT-3Ј; (Cys 18 -Ala) 5Ј-CGCGAGCTGCTCAGCGGTACTCTCATT-3Ј; (Cys 138 -Ala) 5Ј-CAGCCGGGAGGTGGCGTTCTTGTCCTT-3Ј; (Cys 232 -Ala) 5Ј-T-CCCATCAGGGAGGCGCCAGTCTTGGA-3Ј; (Cys 308 -Ala) 5Ј-GATCAG-CAGCAGGGCGGCGCTCTCAGT-3Ј.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Role of Cysteine Residues in Human Angiotensinogen

Gimenez-Roqueplo

Célérier

Schmid

et al. 1998

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

The M235T polymorphism of human angiotensinogen is associated with essential and pregnancy-induced hypertension. A covalent complex is formed between angiotensinogen and the proform of the eosinophil major basic protein (proMBP) during pregnancy. Angiotensinogen is the specific substrate of renin in the renin-angiotensin system. Angiotensinogen belongs to the serine protease inhibitor (serpin) superfamily (1), although it has probably lost this activity (2). Linkage and case-control studies have shown the potential role of the human angiotensinogen gene (hAGT) 1 as a susceptibility gene to develop hypertension (reviewed in Ref.3). The M235T angiotensinogen polymorphism, which is in strong linkage disequilibrium with the G-6A substitution in the promoter (4), has been associated with essential (5) and pregnancy-induced (6) hypertension.Purified plasma angiotensinogen is a heterogeneous protein which migrates at around 60 kDa on SDS-polyacrylamide gel electrophoresis (SDS-PAGE). In addition to this classical monomeric form of angiotensinogen, a high molecular weight form (HMW) has been described in human species (Ref. 7; for a review, see Ref. 8) which is a major component of the placentaspecific renin-angiotensin system, predominant in amniotic fluid (9, 10) and in extracts of placental tissue (11). The plasma of men and menstruating women contains around 5% of the total angiotensinogen as HMW angiotensinogen (12-14) and this can reach 9.6% in women on estrogen contraceptive (12). The percentage of the HMW form increases to 16% in normal pregnancy, and can reach 25-28% in pregnancy-induced hypertension (12). Kinetic studies have shown that renin produces angiotensin I (Ang I) 1.9 times faster from monomeric than from HMW angiotensinogen at physiological pH (15). Previous studies using disulfide bridge reduction (16) and immunoreactivity analysis (17) have suggested that the HMW angiotensinogen could be a covalent complex of monomeric angiotensinogen with other angiotensinogen molecules, or other proteins. Recently, the purification of the proform of the human eosinophil major basic protein (proMBP) from the serum of pregnant women revealed two circulating heterodimeric complexes of angiotensinogen and proMBP covalently linked by disulfide bonds. These complexes were a 2:2 complex of angiotensinogen and proMBP, or a 2:2:2 complex of angiotensinogen, proMBP and complement C3dg (18). They could contribute, at least in part, to HMW angiotensinogen.The sequence of human angiotensinogen contains four cysteine residues at amino acid positions 18, 138, 232, and 308. Only the Cys 18 and Cys 138 are present in all four mammalian species cloned so far (19 -22). Nothing is known about the possible presence of a disulfide bond in angiotensinogen, and consequently which cysteine could interact with proMBP. This study was done to identify cysteine residue of human angiotensinogen involved in a disulfide bridge with proMBP and to study the formation and the function of the angiotensinogen⅐proMBP complex in vitro. We carried out ...

show abstract

Role of N-Glycosylation in Human Angiotensinogen

Cited by 38 publications

References 42 publications

Antinociceptive Structure-Activity Studies with Enkephalin-Based Opioid Glycopeptides

Antinociceptive Structure-Activity Studies with Enkephalin-Based Opioid Glycopeptides

Ser84 of Human Renin Contributes to the Biphasic pH Dependence of the Renin-Angiotensinogen Reaction

Role of Cysteine Residues in Human Angiotensinogen

Contact Info

Product

Resources

About