scholarly journals Identification of a second neutrophil-chemoattractant cytokine generated during an inflammatory reaction in the rabbit peritoneal cavity in vivo. Purification, partial amino acid sequence and structural relationship to melanoma-growth-stimulatory activity

1991 ◽  
Vol 278 (2) ◽  
pp. 493-497 ◽  
Author(s):  
P J Jose ◽  
P D Collins ◽  
J A Perkins ◽  
B C Beaubien ◽  
N F Totty ◽  
...  
Keyword(s):  
Amino Acid ◽  
Cation Exchange ◽  
Reversed Phase ◽  
Interleukin 8 ◽  
Amino Acid Sequencing ◽  
Terminal Amino Acid ◽  
Stimulatory Activity ◽  
Terminal Amino ◽  
Melanoma Growth

The intraperitoneal injection of zymosan in the rabbit results in the generation of an inflammatory exudate containing oedema-forming and chemoattractant activities. Previous studies demonstrated the early appearance of the complement fragment C5a, followed by the generation of two mediators related to the cytokine interleukin-8 that were separable by cation-exchange h.p.l.c. N-Terminal amino acid sequencing identified one of these mediators as rabbit interleukin-8. This paper describes the purification of the second cytokine by cation-exchange, gel-filtration and reversed-phase h.p.l.c. The purified material had both oedema-forming and chemoattractant activity when assayed in rabbit skin in vivo. On SDS/PAGE a single 6-8 kDa band was observed and N-terminal amino acid sequencing of the reduced and alkylated protein positively identified 36 amino acids. This sequence revealed the rabbit homologue of melanoma-growth-stimulatory activity. The identification of these two cytokines in vivo will provide an opportunity to investigate the importance of their co-release in the inflammatory process.

scholarly journals A synthetic glucagon-like peptide-1 analog with improved plasma stability

1998 ◽  
Vol 159 (1) ◽  
pp. 93-102 ◽  
Author(s):  
U Ritzel ◽  
U Leonhardt ◽  
M Ottleben ◽  
A Ruhmann ◽  
K Eckart ◽  
...  
Keyword(s):  
Amino Acid ◽  
Plasma Insulin ◽  
Rat Plasma ◽  
Plasma Stability ◽  
Terminal Amino Acid ◽  
Terminal Amino ◽  
Glucagon Like Peptide ◽  
Glucagon Like Peptide 1

Glucagon-like peptide-1 (GLP-1) is the most potent endogenous insulin-stimulating hormone. In the present study the plasma stability and biological activity of a GLP-1 analog, [Ser]GLP-1(7-36)amide, in which the second N-terminal amino acid alanine was replaced by serine, was evaluated in vitro and in vivo. Incubation of GLP-1 with human or rat plasma resulted in degradation of native GLP-1(7-36)amide to GLP-1(9-36)amide, while [Ser]GLP-1(7-36)amide was not significantly degraded by plasma enzymes. Using glucose-responsive HIT-T15 cells, [Ser]GLP-1(7-36)amide showed strong insulinotropic activity, which was inhibited by the specific GLP-1 receptor antagonist exendin-4(9-39)amide. Simultaneous i.v. injection of [Ser]GLP-1(7-36)amide and glucose in rats induced a twofold higher increase in plasma insulin levels than unmodified GLP-1(7-36)amide with glucose and a fivefold higher increase than glucose alone. [Ser]GLP-1(7-36)amide induced a 1.5-fold higher increase in plasma insulin than GLP-1(7-36)amide when given 1 h before i.v. application of glucose. The insulinotropic effect of [Ser]GLP-1(7-36)amide was suppressed by i.v. application of exendin-4(9-39)amide. The present data demonstrate that replacement of the second N-terminal amino acid alanine by serine improves the plasma stability of GLP-1(7-36)amide. The insulinotropic action in vitro and in vivo was not impaired significantly by this modification.

High Expression of Human Amelogenin in E. Coli

1996 ◽  
Vol 10 (2) ◽  
pp. 187-194 ◽  
Author(s):  
D. Deutsch ◽  
E. Chityat ◽  
M. Hekmati ◽  
A. Palmon ◽  
Y. Farkash ◽  
...  
Keyword(s):  
Amino Acid ◽  
Western Blotting ◽  
Rt Pcr ◽  
Amino Acid Sequencing ◽  
Terminal Amino Acid ◽  
Expression Plasmid ◽  
Over Expression ◽  
E Coli ◽  
Sds Page ◽  
Terminal Amino

A human cDNA, encoding for the 175-aminoacid human amelogenin, was prepared by RT PCR from tooth bud mRNA and sub-cloned into pGEX-KG expression plasmid for over-expression in E. coli. The expressed protein was characterized by SDS-PAGE, Western blotting, and N-terminal amino acid sequencing.

A LOW MOLECULAR WEIGHT ENDOMETRIAL SECRETORY PROTEIN WHICH IS INCREASED BY OVINE TROPHOBLAST PROTEIN-1 IS A β2-MICROGLOBULIN-LIKE PROTEIN

1991 ◽  
Vol 130 (2) ◽  
pp. R1-R4 ◽  
Author(s):  
J. L. Vallet ◽  
P. J. Barker ◽  
G. E. Lamming ◽  
N. Skinner ◽  
N. S. Huskisson
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Secretory Protein ◽  
Explant Culture ◽  
Low Molecular Weight ◽  
Amino Acid Sequencing ◽  
Terminal Amino Acid ◽  
Similar Molecular Weight ◽  
Β2 Microglobulin ◽  
Terminal Amino

ABSTRACT Ovine trophoblast protein-1 (oTP-1), stimulates the secretion of several proteins in explant culture of day-12 cyclic ovine endometrium. We partially purified and identified one of these proteins, an 11,000 Mr, pI approx. 6 protein by N-terminal amino acid sequencing and immunoprecipitation using antibody to human β2-microglobulin. The protein was purified from cultures of endometrium collected from day-16 pregnant ewes. The N-terminal amino acid sequence was 40–55% homologous to β2-microglobulin from a variety of species. Antibody to human β2-microglobulin immunoprecipitated the protein and another protein of similar molecular weight but more acidic pi. Using immunoprecipitation of radiolabelled proteins from culture, we demonstrated that oTP-1 increased production of this protein by 40% (P<0.05). We conclude that oTP-1 increases the secretion of a β2-microglobulin-like protein from day-12 non-pregnant endometrium in culture.

Intramolecular mapping of Plasmodium falciparum P126 proteolytic fragments by N-terminal amino acid sequencing

1992 ◽  
Vol 53 (1-2) ◽  
pp. 89-95 ◽  
Author(s):  
Alain Debrabant ◽  
Pierrette Maes ◽  
Patrick Delplace ◽  
Jean-François Dubremetz ◽  
André Tartar ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Amino Acid ◽  
Amino Acid Sequencing ◽  
Terminal Amino Acid ◽  
Terminal Amino

scholarly journals Evaluation of Human Serum Albumin Cobalt Binding Assay for the Assessment of Myocardial Ischemia and Myocardial Infarction

2003 ◽  
Vol 49 (4) ◽  
pp. 581-585 ◽  
Author(s):  
Nadhipuram V Bhagavan ◽  
Ernest M Lai ◽  
Patricia A Rios ◽  
Jinsheng Yang ◽  
Anna M Ortega-Lopez ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Amino Acid ◽  
Myocardial Ischemia ◽  
Roc Curve ◽  
Binding Assay ◽  
Albumin Binding ◽  
Amino Acid Residues ◽  
Amino Acid Sequencing ◽  
Terminal Amino Acid ◽  
Terminal Amino

Abstract Background: Clinical diagnoses were correlated with results of a Co(II)–albumin binding assay in 167 patients treated at an emergency department of a health maintenance organization. Methods: Patients were evaluated as being nonischemic or potentially ischemic through standard coronary disease indicators [creatine kinase (CK), CK-MB, cardiac troponin I, and electrocardiographic findings] and were tested by a Co(II)–albumin binding assay. Samples were tested anonymously, and the study was double-blinded. The sensitivity and specificity of this assay for the detection of ischemia were evaluated by ROC curve analysis. Known Co(II) binding sites on albumin were analyzed by N-terminal amino acid sequencing. Results: The mean absorbance units (ABSU) ± 2 SD for non-myocardial ischemic and myocardial ischemic individuals measured at 470 nm were 0.43 ± 0.10 and 0.63 ± 0.25, respectively (P &lt;0.0001). The area under the ROC curve was 0.95 [95% confidence interval (CI), 0.92–0.99], and at a cutoff value of 0.50 ABSU, sensitivity and specificity were 88% (78–94%) and 94% (86–98%), respectively, suggesting a high distinction between the two groups. When we compared non-acute myocardial infarction (AMI) and AMI ischemic individuals, the area under the ROC curve was 0.66 (95% CI, 0.53–0.79) and was considered a poor discriminator between these two groups. N-Terminal amino acid sequencing data for purified albumin showed normal amino acid residues for six of seven high-ABSU (≥0.70) individuals and one nonischemic individual tested. However, only one individual with a high ABSU (0.80) had two missing amino acid residues (DA) from the N-terminal region. Clinical diagnosis for this patient did not reveal an ischemic event. Conclusions: The Co(II)–albumin binding test may serve as a useful diagnostic tool in emergency facilities for the assessment of myocardial ischemia. High and low ABSU were associated with myocardial ischemic individuals and non-myocardial ischemic individuals, respectively. However, the Co(II)–albumin binding was a poor discriminator between ischemic individuals with and without MI.

Partial Purification and N-Terminal Amino Acid Sequencing of a β-1,3-Glucanase from Sorghum Leaves

2003 ◽  
Vol 46 (1) ◽  
pp. 29-33 ◽  
Author(s):  
R. Velazhahan ◽  
J. Jayaraj ◽  
G.H. Liang ◽  
S. Muthukrishnan
Keyword(s):  
Amino Acid ◽  
Partial Purification ◽  
Amino Acid Sequencing ◽  
Terminal Amino Acid ◽  
Terminal Amino
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