scholarly journals The behaviour of peptides on reverse-phase supports during high-pressure liquid chromatography

1981 ◽  
Vol 199 (1) ◽  
pp. 31-41 ◽  
Author(s):  
K J Wilson ◽  
A Honegger ◽  
R P Stötzel ◽  
G J Hughes
Keyword(s):  
High Pressure ◽  
Liquid Chromatography ◽  
Amino Acid ◽  
Chain Length ◽  
Solid Phase ◽  
Chromatographic Retention ◽  
Buffer System ◽  
Amino Acid Residues ◽  
Reverse Phase ◽  
Chromatographic Behaviour

High-pressure (‘performance’) liquid chromatography has been used to investigate the reverse-phase chromatographic behaviour of peptides, ranging in length from 2 to 65 amino acid residues, which have originated from primary-sequence determinations or solution/solid-phase syntheses. By using a pyridine/formate-pyridine/acetate/propan-1-ol buffer system, as previously described [Hughes, Winterhalter & Wilson (1979) FEBS Lett. 108, 81-86], the influence of various experimental parameters were examined. (a) Peptide retention was observed to be temperature-independent between 25 and 55 degrees C. (b) The dependence of chromatographic retention on pH decreases with increasing peptide hydrophobicity. (c) Chromatographic results from C8- and C18-chain-length, as well as from 5 micrometers- and 10 micrometers-particle-size, supports were comparable. (d) The hydrophobic strength of the organic solvent in the mobile phase was observed to decrease: propan-1-ol approximately equal to propan-2-ol greater than acetonitrile much greater than methanol. (e) When gradient rates (% of buffer B/unit time) were systematically decreased, peptide retention decreased in a hyperbolic manner. Comparisons of the peptides chromatographed with respect to their measured retention properties and calculated hydrophobicities were performed by computer analysis. Deviation of peptide chromatographic behaviour was observed to be essentially independent of hydrophobicity, chain length and charge. On the basis of the measured retention properties of the chromatographed peptides, hydrophobic constants for the various amino acid side chains were determined and compared with similar constants available from the literature.

scholarly journals Comparison of the primary structures of ribonuclease U2 isoforms

1986 ◽  
Vol 240 (1) ◽  
pp. 163-170 ◽  
Author(s):  
S Kanaya ◽  
T Uchida
Keyword(s):  
Amino Acid ◽  
Carboxy Group ◽  
Side Chain ◽  
Amino Acid Residues ◽  
Chemical Analyses ◽  
Reverse Phase ◽  
Chromatographic Behaviour ◽  
Isopeptide Bond ◽  
Primary Structures ◽  
Asparagine Residue

The primary structures of the two isoforms of ribonuclease U2, RNAases U2-A and U2-B, were analysed and compared with each other. Among the chymotryptic peptides obtained from the reduced and S-carboxymethylated enzymes, only peptides C-3 were different from each other in terms of chromatographic behaviour on reverse-phase h.p.l.c. On the basis of chemical analyses of these peptides, it was shown that RNAase U2-B had an isopeptide bond in which Asp-32 was linked to Gly-33 through the beta-carboxy group in its side chain instead of the alpha-carboxy group. Deamidation of Asn-32 in RNAase U2-A led to the formation of this unusual linkage. The previously reported sequence of RNAase U2 [Sato & Uchida (1975) Biochem. J. 145, 353-360] was corrected by changing amino acid residues at eight different positions and by inserting an asparagine residue at position 32. The numbering of the positions of amino acid residues located downstream of Asn-32 was therefore shifted by 1. Accordingly, RNAase U2-A was shown to be composed of 114 amino acid residues.

scholarly journals Microcin E492 Is an Unmodified Peptide Related in Structure to Colicin V

2002 ◽  
Vol 46 (1) ◽  
pp. 229-230 ◽  
Author(s):  
Anne-Marie Pons ◽  
Nathalie Zorn ◽  
David Vignon ◽  
François Delalande ◽  
Alain Van Dorsselaer ◽  
...  
Keyword(s):  
High Pressure ◽  
Liquid Chromatography ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Solid Phase Extraction ◽  
Molecular Mass ◽  
Solid Phase ◽  
Reversed Phase ◽  
Phase Extraction ◽  
Colicin V

ABSTRACT The pore-forming microcin E492 was purified by solid-phase extraction and reversed-phase high-pressure liquid chromatography. Its molecular mass was 7,886 Da. The entire 84-amino-acid sequence was determined. There is no postranslational modification in the secreted microcin, and the sequence has homologies with the sequence of the microcin colicin V.

Oxytocin analogues with non-coded amino acid residues in position 8: [8-Neopentylglycine]oxytocin and [8-cycloleucine]oxytocin

1987 ◽  
Vol 52 (9) ◽  
pp. 2317-2325 ◽  
Author(s):  
Jan Hlaváček ◽  
Jan Pospíšek ◽  
Jiřina Slaninová ◽  
Walter Y. Chan ◽  
Victor J. Hruby
Keyword(s):  
Amino Acid ◽  
Solid Phase Synthesis ◽  
Solid Phase ◽  
The Other ◽  
Amino Acid Residues ◽  
Phase Synthesis ◽  
Other Hand ◽  
Fragment Condensation

[8-Neopentylglycine]oxytocin (II) and [8-cycloleucine]oxytocin (III) were prepared by a combination of solid-phase synthesis and fragment condensation. Both analogues exhibited decreased uterotonic potency in vitro, each being about 15-30% that of oxytocin. Analogue II also displayed similarly decreased uterotonic potency in vivo and galactogogic potency. On the other hand, analogue III exhibited almost the same potency as oxytocin in the uterotonic assay in vivo and in the galactogogic assay.

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