Site Directed Antisera to the D-2 Polypeptide Subunit of Photosystem II

Three synthetic oligopeptides were used for preparation of antibodies against the D-2 polypeptide of thylakoid membranes. Their sequence was chosen from a model of the folding of the amino acid sequence of the D-2 polypeptide subunit through the membrane that predicted these sequences to be exposed at the membrane surface. For the Merrifield solid-phase method on a fully automated synthesizer the Na-amino group was protected by a fluorenyl-9-methylcarbonyl group. The oligopeptides were coupled to serum albumin by EDAC for immunizations in rabbits. Antisera with high titer were obtained for the two oligopeptides that contained the amino acid sequence of the D-2 protein from amino acid 230 to 235 and from 235 to 241. The antisera reacted with the D-2 polypeptide, separated on SDS gel and agglutinated the thylakoid membrane. It is known that certain photosystem II functions are impaired by short time trypsin treatment of the membrane. The antisera were used to show that under these conditions the D-2 polypeptide in the membrane is very sensitive. The trypsination yielded two cleavage products detected by the two antisera, a 20 kDa fragment blotted by antiserum against amino acids 230 to 235 and a 10 kDa fragment blotted by the antiserum against amino acids 235 to 241. As the polypeptide cleavage occurs between the two epitopes, the trypsin cut is therefore at arginine 234. This supports the prediction that the sequence containing this arginine is the most exposed part of the D-2 polypeptide on the membrane (matrix) surface. It is proposed that the high sensitivity of the D-2 polypeptide accounts for the known effect of membrane trypsination on QA accessibility in photosystem II.

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Synthesis and biological activity of new metallocenic angiotensin II analogs

Collection of Czechoslovak Chemical Communications ◽

10.1135/cccc19882914 ◽

1988 ◽

Vol 53 (11) ◽

pp. 2914-2919 ◽

Cited By ~ 10

Author(s):

Pierrette Maes ◽

Annie Ricouart ◽

Emmanuel Escher ◽

André Tartar ◽

Christian Sergheraert

Keyword(s):

Amino Acids ◽

Biological Activity ◽

Angiotensin Ii ◽

Solid Phase ◽

Rabbit Aorta ◽

Phase Method ◽

Solid Phase Method

Analogs of angiotensin II in which phenylalanine in position 8 was replaced with cymantrenylalanine or with its triphenylphosphine photosubstitution product were synthesized by the solid-phase method. On rabbit aorta strips, these peptides were found to be pure antagonists of angiotensin II. Their relative affinities are higher than most other analogs substituted in position 8 with bulky amino-acids.

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Combustion Synthesis of La0.65Sr0.35MnO3 and its Sensing Properties for NO2

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.680.208 ◽

2016 ◽

Vol 680 ◽

pp. 208-211

Author(s):

Lian Lian Wu ◽

Qiang Li ◽

Dan Yu Jiang ◽

Jin Feng Xia

Keyword(s):

Solid Phase ◽

High Sensitivity ◽

Combustion Method ◽

Fast Response ◽

Size Analysis ◽

Phase Method ◽

X Ray Diffraction ◽

Ultrafine Structure ◽

Nox Sensor ◽

Solid Phase Method

In this paper, La0.65Sr0.35MnO3 (LSM) oxide powder with ultrafine structure has been synthesized by self-propagating combustion method. The powders were characterized by X-ray diffraction, scanning electron microscopy and laser size analysis. Compared to the powders prepared by traditional solid-phase method, the grain size of powders prepared by self-propagating combustion method is relatively small and uniform. Starting from ultrafine LSM powders, sensing electrode (SE) for NO2 mixed-potential sensors based on yttria-stablized zirconia (YSZ) was fabricated. As-obtained NO2 sensor displays fast response and high sensitivity (25.4mV/decade). The response values of the sensor have good linear relationship with the logarithm of NO2 concentration varying from 30ppm to 500ppm.Keywords:Self-propagating combustion method; La0.65Sr0.35MnO3; NOx sensor; YSZ

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Isolation and Characterization of a Type I Gene Encoding a Light-harvesting Chlorophyll a/b-binding Protein of Photosystem II in Peach

Journal of the American Society for Horticultural Science ◽

10.21273/jashs.123.4.493 ◽

1998 ◽

Vol 123 (4) ◽

pp. 493-499 ◽

Cited By ~ 1

Author(s):

Kyu H. Chung ◽

Dennis E. Buetow ◽

Schuyler S. Korban

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Photosystem Ii ◽

Amino Acid Sequence ◽

Woody Plants ◽

Light Harvesting ◽

Binding Protein ◽

Transit Peptide ◽

Type I ◽

Polyadenylation Sites

A nuclear gene, Lhcb1*Pp1, encoding a light-harvesting chlorophyll a/b-binding protein of photosystem II has been isolated from peach [Prunus persica (L.) Batsch. `Stark Earliglo'] leaf genomic DNA, cloned, and sequenced. This gene encodes a precursor polypeptide of 267 amino acids with a transit peptide of 34 and a type I mature protein of 233 amino acids. The amino acid sequence of the mature polypeptide is 89% to 94% and 80% to 94% similar to those encoded by type I Lhcb genes of annual and other woody plants, respectively. In contrast, the amino acid sequence of the peach transit peptide is less conserved being 47% to 69% similar to those of annual plants and only 17% to 22% similar to those of other woody plants. The peach gene was used as a probe for Lhcb gene expression. Lhcb mRNA is detected in leaves of field-grown trees during June to October. Lhcb mRNA is detected at a high level in leaves of peach shoots grown in tissue culture in the light, but only at a trace level in leaves grown in the dark. Some Lhcb genes appear to be light-modulated in stems. Lhcb1*Ppl contains four potential polyadenylation sites. S1 nuclease analysis detected transcripts of the sizes expected from each of the four polyadenylation sites. All four are found in leaves of light-grown shoots and of field-grown trees throughout the growing season. In contrast, only three are detected in stems of light-grown shoots.

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Synthese eines Thymosin β10-Fragments zur Entwicklung spezifischer Antikörper / Synthesis of a Fragment of Thymosin β10 for the Development of Specific Antibodies

Zeitschrift für Naturforschung B ◽

10.1515/znb-1992-0819 ◽

1992 ◽

Vol 47 (8) ◽

pp. 1170-1174 ◽

Cited By ~ 4

Author(s):

Susanne Hörger ◽

Brigitte Gallert ◽

Hartmut Echner ◽

Wolfgang Voelter

Keyword(s):

Amino Acids ◽

Solid Phase ◽

Protecting Groups ◽

Side Chain ◽

Phase Method ◽

Preparative Hplc ◽

Specific Antibodies ◽

Tert Butyl ◽

Thymosin Β10 ◽

Solid Phase Method

The N-terminal fragment 1-12 of thymosin β10 was synthesized by the solid phase method using p-benzyloxybenzyl alcohol/polystyrene/divinylbenzeneresin and N-a-Fmoc amino acids with tert-butyl or Boc side chain protecting groups. Coupling was performed with BOP. The peptide was purified by preparative HPLC.

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The Dual Regulatory Role of Amino Acids Leu480 and Gln481 of Prothrombin

Journal of Biological Chemistry ◽

10.1074/jbc.m115.691956 ◽

2015 ◽

Vol 291 (4) ◽

pp. 1565-1581 ◽

Cited By ~ 1

Author(s):

Joesph R. Wiencek ◽

Jamila Hirbawi ◽

Vivien C. Yee ◽

Michael Kalafatis

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Amino Acid Sequence ◽

Protein C ◽

Membrane Surface ◽

Point Mutations ◽

Regulatory Subunit ◽

Activate Factor ◽

Factor V ◽

Divalent Metal Ions

Prothrombin (FII) is activated to α-thrombin (IIa) by prothrombinase. Prothrombinase is composed of a catalytic subunit, factor Xa (fXa), and a regulatory subunit, factor Va (fVa), assembled on a membrane surface in the presence of divalent metal ions. We constructed, expressed, and purified several mutated recombinant FII (rFII) molecules within the previously determined fVa-dependent binding site for fXa (amino acid region 473–487 of FII). rFII molecules bearing overlapping deletions within this significant region first established the minimal stretch of amino acids required for the fVa-dependent recognition exosite for fXa in prothrombinase within the amino acid sequence Ser478–Val479–Leu480–Gln481–Val482. Single, double, and triple point mutations within this stretch of rFII allowed for the identification of Leu480 and Gln481 as the two essential amino acids responsible for the enhanced activation of FII by prothrombinase. Unanticipated results demonstrated that although recombinant wild type α-thrombin and rIIaS478A were able to induce clotting and activate factor V and factor VIII with rates similar to the plasma-derived molecule, rIIaSLQ→AAA with mutations S478A/L480A/Q481A was deficient in clotting activity and unable to efficiently activate the pro-cofactors. This molecule was also impaired in protein C activation. Similar results were obtained with rIIaΔSLQ (where rIIaΔSLQ is recombinant human α-thrombin with amino acids Ser478/Leu480/Gln481 deleted). These data provide new evidence demonstrating that amino acid sequence Leu480–Gln481: 1) is crucial for proper recognition of the fVa-dependent site(s) for fXa within prothrombinase on FII, required for efficient initial cleavage of FII at Arg320; and 2) is compulsory for appropriate tethering of fV, fVIII, and protein C required for their timely activation by IIa.

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Antagonistic Analogs of Oxytocin with Substituted Phenylalanine or Tyrosine in Position 2

Collection of Czechoslovak Chemical Communications ◽

10.1135/cccc19941430 ◽

1994 ◽

Vol 59 (6) ◽

pp. 1430-1438 ◽

Cited By ~ 10

Author(s):

Rudolf Ježek ◽

Miroslava Žertová ◽

Jiřina Slaninová ◽

Pavel Majer ◽

Zdenko Procházka

Keyword(s):

Amino Acids ◽

Solid Phase ◽

Phase Method ◽

Inhibiting Activity ◽

Structure Activity ◽

Low Degree ◽

Solid Phase Method ◽

Pressor Activity ◽

Agonistic Activity

Solid phase method on p-methylbenzhydrylamine resin was used for the synthesis of seven analogs of oxytocin with non-coded amino acids in position 2. [L-Phe(4-Me)2]oxytocin (I), [D-Phe(4-Me)2]oxytocin (II), [L-Phe(2-Me,4-Et)2]oxytocin (III), [D-Phe(2-Me,4-Et)2]oxytocin (IV), [D-Tyr(Me)2]oxytocin (V), [D-Tyr(Et)2]oxytocin (VI) and [L-Tyr(2-Me)2]oxytocin (VII) were synthesized. All analogs with D-stereoisomer of alkyl or alkoxy substituted phenylalanine possess uterus in vitro inhibiting activity. In the case of L-stereoisomers the structure activity relationship is more complicated. As far as the pressor activity is concerned, the analogs have either very low agonistic activity or low degree of antagonism.

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The Analogs of [D-Har8]Vasopressin with Di- and Trisubstituted Phenylalanine in Position 2; Synthesis and Some Biological Properties

Collection of Czechoslovak Chemical Communications ◽

10.1135/cccc19932751 ◽

1993 ◽

Vol 58 (11) ◽

pp. 2751-2760 ◽

Cited By ~ 4

Author(s):

Miroslava Žertová ◽

Zdenko Procházka ◽

Jiřina Slaninová ◽

Tomislav Barth ◽

Pavel Majer ◽

...

Keyword(s):

Amino Acids ◽

Solid Phase ◽

Biological Properties ◽

Phase Method ◽

Antidiuretic Activity ◽

Solid Phase Method

Four analogs of vasopressin with non-coded amino acids D-homoarginine (in position 8) and 2,6-di- or 2,4,6-trisubstituted L- or D-phenylalanine (in position 2) were synthesized using the solid phase method on p-methylbenzhydrylamine resin. All the analogs were found to be uterotonic inhibitors, the most potent one in vitro and in vivo being [D-Phe(2,4,6-triMe)2,D-Har8]vasopressin with pA2 values equal to 8.1 and 7.5, respectively. All of them had negligible antidiuretic activity and were weak pressor inhibitors.

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New analogs of arginine-vasopressin containing β-homo-L-amino acid residues

Collection of Czechoslovak Chemical Communications ◽

10.1135/cccc19892795 ◽

1989 ◽

Vol 54 (10) ◽

pp. 2795-2801 ◽

Cited By ~ 2

Author(s):

Krysztof Bankowski ◽

Alexandra Misicka ◽

Tomislav Barth ◽

Jiřina Slaninová

Keyword(s):

Amino Acid ◽

Amino Acid Residue ◽

Arginine Vasopressin ◽

Solid Phase ◽

Phase Method ◽

Amino Acid Residues ◽

Solid Phase Method

Four new analogs of arginine vasopressin containing β-homo-L-amino acid residue were synthesized by the solid-phase method. The introduced modifications yielded the following peptides:[β-homo Phe3]AVP (I), [β-homoPro7]AVP (II), [Cpp1, Tyr(Me)2, β-homoPhe3]AVP (III), and[Cpp1,Tyr(Me)2, β-homoPro7]AVP (IV). Agonistic properties of I and II, as well as antagonistic properties of III and IV were decreased, more pronouncedly with analogs substituted in the position 3.

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A simple and traceless solid phase method simplifies the assembly of large peptides and the access to challenging proteins

Chemical Science ◽

10.1039/c7sc01912b ◽

2017 ◽

Vol 8 (8) ◽

pp. 5362-5370 ◽

Cited By ~ 14

Author(s):

N. Ollivier ◽

R. Desmet ◽

H. Drobecq ◽

A. Blanpain ◽

E. Boll ◽

...

Keyword(s):

Amino Acids ◽

Solid Phase ◽

Biologically Active ◽

Phase Method ◽

Active Protein ◽

Solid Phase Method

We show that the combination of solid phase and solution ligation techniques facilitates the production of a challenging and biologically active protein made of 180 amino acids.

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