scholarly journals Rapid purification and properties of potassium-activated aldehyde dehydrogenase from Saccharomyces cerevisiae

1978 ◽  
Vol 173 (3) ◽  
pp. 773-786 ◽  
Author(s):  
K A Bostian ◽  
G F Betts
Keyword(s):  
Amino Acid ◽  
Aldehyde Dehydrogenase ◽  
Amino Acid Content ◽  
Binding Studies ◽  
Terminal Amino Acid ◽  
Purification And Properties ◽  
Direct Binding ◽  
Terminal Amino ◽  
Rapid Purification

A method for the purification of yeast K+-activated aldehyde dehydrogenase is presented which can be completed in substantially less time than other published procedures. The enzyme has a different N-terminal amino acid from preparations previously reported, and other small differences in amino acid content. These differences may be the result of differential proteolytic digestion rather than a different protein in vivo. A purification step involves the biospecific adsorption on affinity columns containing immobilized nucleotides in the absence of the substrate aldehyde. Direct binding studies with the coenzyme in the absence of aldehyde reveal 4 NAD sites per tetrameric molecule, each with a dissociation constant of 120 micron. These results conflict with properties of preparations previously reported and may conflict with kinetic models that have aldehyde as the leading substrate. Binding to Blue Dextran affinity columns suggests the presence of a dinucleotide fold in common with other dehydrogenases and kinases.

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.

scholarly journals Acetylcholinesterase aus Rindererythrozyten. Reinigung und Eigenschaften des mit und ohne Triton X-100 solubilisierten Enzyms / Acetylcholinesterase from Bovine Erythrocytes. Purification and Properties of the En­zyme Solubilized in the Presence and the Absence of Triton X-100

1979 ◽  
Vol 34 (9-10) ◽  
pp. 721-725 ◽  
Author(s):  
Heinz Großmann ◽  
Manfred Liefländer
Keyword(s):  
Amino Acid ◽  
Specific Activity ◽  
Multiple Forms ◽  
Terminal Amino Acid ◽  
Enzyme Preparations ◽  
Purification And Properties ◽  
Solubilized Enzyme ◽  
Terminal Amino ◽  
Triton X ◽  
Bovine Erythrocytes

Abstract Acetylcholinesterase was released from bovine erythrocytes by Triton X-100 treatment and pu­rified by twofold affinity chromatography. The detergentfree enzyme was obtained with a specific activity of 4130 U /mg (303 000-fold purification) and a 25% yield. Alternatively, the commercial available crude enzyme was purified. The latter preparation has an uniform molecular weight (Mr 175 000). The Triton-solubilized enzyme, however, can be resolved after removal of the detergent in eight multiple forms (Mr 175 000 and multiple values), in the presence of Triton there exists only one form (Mr 338 000). The amino acid composition of the two enzyme preparations differs significantly. No differences were observed with respect to other properties: SDS gel electrophore­sis revealed two protein bands (Mr 166 000 and 86 000) with both preparations. The enzyme is a glycoprotein with a pI value of 4.3 and contains strongly bound phosphatidylethanolamine. The N-terminal amino acid has been found to be Glu (or Gin).

scholarly journals Purification and Properties of Staphylocoagulase

1975 ◽  
Author(s):  
A.D. Muller ◽  
B. M. Bas ◽  
H. C. Hemker
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Amino Acid Composition ◽  
Aspartic Acid ◽  
Acid Composition ◽  
Isoelectric Point ◽  
Terminal Amino Acid ◽  
Purification And Properties ◽  
Terminal Amino

Staphylocoagulase, an exoprotein of coagulase positive staphylocoagulase, has been purified to a state in which only trace amounts of contaminating proteins are detectable.Purification was more than 35,000 fold, which is 7 times more than the highest value reported in the literature. The yield was about 15%.Aspartic acid was found as a single N-terminal amino acid in this preparation. The molecular weight is 61,000 and the isoelectric point lies at pH 4.53.The amino acid composition was determined.

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.

Purification and Properties of an Enzyme Capable of Degrading the Polysaccharide of the Cyanobacterium, Nostoc commune

2002 ◽  
Vol 57 (11-12) ◽  
pp. 1042-1046 ◽  
Author(s):  
◽  
Shin’ichiro Kajiyama ◽  
Atsushi Okazawa ◽  
Ei-ichiro Fukusaki ◽  
Akio Kobayashi
Keyword(s):  
Enzyme Activity ◽  
Amino Acid ◽  
Nostoc Commune ◽  
Terminal Amino Acid ◽  
Degrading Enzyme ◽  
Purification And Properties ◽  
Optimum Ph ◽  
Terminal Amino ◽  
Terminal Amino Acid Sequence ◽  
Optimum Ph And Temperature

A novel Nostoc commune-polysaccharide (NPS)-degrading enzyme with a molecular mass of 128.5 kDa was purified from Paenibacillus glycanilyticus DS-1. The optimum pH and temperature of the enzyme activity were 5.5 and 35 °C, respectively. The enzyme completely degraded NPS to oligosaccharides, ranging from tetra to hexasaccharides and could degrade the xylan weakly whereas xanthan, gellan, cellulose, curdlan and p-nitrophenyl-β-ᴅ-xylopyranoside were not degraded. Homology analysis of the N-terminal amino acid sequence of the NPS-degrading enzyme against the PIR and SWISS-PROT databases indicated that the sequence was not homologous to any other polysaccharide-degrading enzyme.

Acetylcholinesterase aus dem Gift von Bungarus multicinctus. Reinigung und Eigenschaften/The Acetylcholinesterase of Bungarus multicinctus Venom. Purification and Properties

1979 ◽  
Vol 34 (1-2) ◽  
pp. 27-32 ◽  
Author(s):  
H. Großmann ◽  
M. Weinert ◽  
M. Liefländer
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Isoelectric Focusing ◽  
Gel Electrophoresis ◽  
Amino Acid Analysis ◽  
Specific Activity ◽  
Terminal Amino Acid ◽  
Purification And Properties ◽  
Gradient Gel Electrophoresis ◽  
Terminal Amino

Abstract Acetylcholinesterase from Banded krait (Bungarus multicinctus) venom has been purified by CM-Sephadex chromatography and affinity chromatography to a specific activity of 4290 U/mg. The purified enzyme is a glycoprotein. It is free of electrophoretically detectable contaminating proteins. A molecular weight of 140 000 ± 5 000 has been determined by gradient gel electrophoresis for the native enzyme. It is split into two equal-sized subunits (Mr 70 000 ± 2 000) by SDS treatment. The N-terminal amino acid analysis gave glycine and serine. The purified acetyl­ cholinesterase can be resolved by disc gel electrophoresis into four and by isoelectric focusing into six isozymes. The pI value of the main isozyme has been found to be 5.98 ± 0.05.

Global in vivo terminal amino acid labeling for exploring differential expressed proteins induced by dialyzed serum cultivation

The Analyst ◽  
2014 ◽  
Vol 139 (18) ◽  
pp. 4497-4504 ◽  
Author(s):  
Li-Qi Xie ◽  
Ai-Ying Nie ◽  
Shu-Jun Yang ◽  
Chao Zhao ◽  
Lei Zhang ◽  
...  
Keyword(s):  
Amino Acid ◽  
High Throughput ◽  
Trypsin Digestion ◽  
Metabolic Labeling ◽  
Terminal Amino Acid ◽  
Terminal Amino ◽  
Amino Acid Labeling ◽  
Dialyzed Serum

An accurate and high throughput isobaric MS2 quantification strategy based on metabolic labeling and trypsin digestion.

Fractionation of hordein by preparative, continuous carrier-free electrophoresis

1968 ◽  
Vol 46 (10) ◽  
pp. 1301-1307 ◽  
Author(s):  
Ch. Ivanov ◽  
B. Mesrob ◽  
Z. Prusik
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Acid Content ◽  
Amino Acid Content ◽  
Amino Acid Residues ◽  
Terminal Amino Acid ◽  
Basic Fraction ◽  
Acidic Amino Acids ◽  
Carrier Free ◽  
Terminal Amino

Barley hordein was fractionated by preparative, continuous carrier-free electrophoresis. Six fractions were obtained, one of which was in negligible quantity. Three of the fractions gave single symmetrical peaks. The amino acid content and the N-terminal amino acid residues of these fractions were determined. The ratios of basic to acidic amino acids showed that the fractions contained different protein substances. The most basic fraction, representing 10% of the total hordein, appeared to be pure since it contained only alanine as a N-terminal amino acid.

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