Journal of chromatography最新文献_第5页

High-performance liquid chromatography of amino acids, peptides and proteins. CXXVIII. Effect of D-amino acid substitutions on the reversed-phase high-performance liquid chromatography retention behaviour of neuropeptide Y[18-36] analogues. 氨基酸、多肽和蛋白质的高效液相色谱。CXXVIII。d-氨基酸取代对神经肽Y[18-36]类似物反相高效液相色谱保留行为的影响。

Journal of chromatography Pub Date : 1993-08-27 DOI: 10.1016/s0021-9673(99)87007-1

M I Aguilar, S Mougos, J Boublik, J Rivier, M T Hearn

{"title":"High-performance liquid chromatography of amino acids, peptides and proteins. CXXVIII. Effect of D-amino acid substitutions on the reversed-phase high-performance liquid chromatography retention behaviour of neuropeptide Y[18-36] analogues.","authors":"M I Aguilar, S Mougos, J Boublik, J Rivier, M T Hearn","doi":"10.1016/s0021-9673(99)87007-1","DOIUrl":"https://doi.org/10.1016/s0021-9673(99)87007-1","url":null,"abstract":"The reversed-phase high-performance liquid chromatographic (RP-HPLC) gradient elution behaviour of a series of peptides related to Neuropeptide Y (NPY) has been investigated. The peptides studied included NPY, NPY[13-36], NPY[18-36] and a series of 16 analogues of NPY[18-36], each with a single D-amino acid substitution. Chromatographic parameters which relate to the interactive contact area and the binding affinity have been evaluated with two different stationary phase ligands and two organic modifiers. The results demonstrate that D-amino acid substitutions in the sequence region encompassing amino acid residues NPY[27-31] of these NPY[18-36] peptides significantly influence the interactive behaviour of these peptides relative to the unsubstituted NPY[18-36] molecule, while substitutions in the N- and C-terminal regions had little effect. Further, these results indicate that, in hydrophobic environments, NPY[18-36] adopts a significant degree of secondary structure which is severely disrupted by the presence of the D-amino acids in the central portion of the molecule.","PeriodicalId":15508,"journal":{"name":"Journal of chromatography","volume":"646 1","pages":"53-65"},"PeriodicalIF":0.0,"publicationDate":"1993-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/s0021-9673(99)87007-1","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19392099","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 42

High-performance liquid chromatography of amino acids, peptides and proteins. CXXVI. Modelling of protein adsorption with non-porous and porous particles in a finite bath. 氨基酸、多肽和蛋白质的高效液相色谱。CXXVI。模拟蛋白质吸附与非多孔和多孔颗粒在有限的浴。

Journal of chromatography Pub Date : 1993-08-27 DOI: 10.1016/s0021-9673(99)87008-3

Q M Mao, R Stockmann, I G Prince, M T Hearn

{"title":"High-performance liquid chromatography of amino acids, peptides and proteins. CXXVI. Modelling of protein adsorption with non-porous and porous particles in a finite bath.","authors":"Q M Mao, R Stockmann, I G Prince, M T Hearn","doi":"10.1016/s0021-9673(99)87008-3","DOIUrl":"https://doi.org/10.1016/s0021-9673(99)87008-3","url":null,"abstract":"Analytical solutions for a mathematical model describing dynamic adsorption processes of proteins onto non-porous adsorbent particles in a finite bath are presented. The model, based on the Langmuir adsorption isotherm, has been applied to experimental data obtained with affinity and ion-exchange adsorbents. The external film mass transfer resistance, as well as the rate of surface interaction between proteins and adsorbents, have been taken into account. The model has been extended to the case of adsorption onto porous particles by employing a linear driving force approximation for describing mass transfer in the pore fluid. This approach enables the derivation of an effective overall liquid phase mass transfer coefficient, permitting subsequent adaptation of the analytical solutions developed for non-porous particles. The evaluation of the effective liquid phase mass transfer coefficients is also described. Examples of a comparison between predicted and experimental dynamic adsorption curves for both dye-affinity and ion-exchange systems are presented. The application of the model for predicting the optimum operating conditions is discussed.","PeriodicalId":15508,"journal":{"name":"Journal of chromatography","volume":"646 1","pages":"67-80"},"PeriodicalIF":0.0,"publicationDate":"1993-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/s0021-9673(99)87008-3","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19392100","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 29

Rapid high-sensitivity peptide mapping by liquid chromatography-mass spectrometry. 液相色谱-质谱联用快速高敏肽图谱分析。

Journal of chromatography Pub Date : 1993-08-27 DOI: 10.1016/s0021-9673(99)87018-6

K Mock, M Hail, I Mylchreest, J Zhou, K Johnson, I Jardine

引用次数: 21

High-performance liquid chromatography of amino acids, peptides and proteins. CXXIX. Ceramic-based particles as chemically stable chromatographic supports. 氨基酸、多肽和蛋白质的高效液相色谱。CXXIX。陶瓷基颗粒作为化学稳定的色谱载体。

Journal of chromatography Pub Date : 1993-08-27 DOI: 10.1016/s0021-9673(99)87014-9

H J Wirth, K O Eriksson, P Holt, M Aguilar, M T Hearn

{"title":"High-performance liquid chromatography of amino acids, peptides and proteins. CXXIX. Ceramic-based particles as chemically stable chromatographic supports.","authors":"H J Wirth, K O Eriksson, P Holt, M Aguilar, M T Hearn","doi":"10.1016/s0021-9673(99)87014-9","DOIUrl":"https://doi.org/10.1016/s0021-9673(99)87014-9","url":null,"abstract":"Porous zirconia based particles have been modified using different derivatisation procedures. The modified particles were characterised in terms of their accessible surface areas and degree of surface coverage of the bounded or physicoated phases utilising the strong and specific adsorption of phosphate ions to the zirconia surface. The hydroxyl group density was determined by a 1H NMR technique. The particles were modified by immobilising different silanes to introduce either hydrophobic ligands or reactive groups onto the zirconia surface. In the latter case, various ligands were then covalently attached to the activated supports. Using this type of modification, n-octadecyl- (C18), carbohydrate- and Cibacron Blue F3GA-modified zirconia particles were produced. Furthermore, polymeric coated particles were prepared either by using polybutadiene or by cross-linking the carbohydrate modified sorbents. The pH stability of the different sorbents were determined in batch experiments and under chromatographic conditions. The leakage of ligands was monitored by UV absorption and by employing radioactively labelled ligands. The performance of the C18 reversed-phase modified zirconia in packed columns was also used as an indicator of changes in the surface chemistry following pH stability tests. The experimental results indicate that the Cibacron Blue F3GA dye-modified sorbent was stable up to pH 10.5, the C18 reversed-phase packing up to pH 13 and the carbohydrate-bonded phase up to pH 12. These investigations substantiate the favourable chemical and physical characteristics anticipated for surface modified zirconias for potential use as chromatographic adsorbents.","PeriodicalId":15508,"journal":{"name":"Journal of chromatography","volume":"646 1","pages":"129-41"},"PeriodicalIF":0.0,"publicationDate":"1993-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/s0021-9673(99)87014-9","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19394027","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 37

Effect of the alpha-amino group on peptide retention behaviour in reversed-phase chromatography. Determination of the pK(a) values of the alpha-amino group of 19 different N-terminal amino acid residues. -氨基对反相色谱中肽保留行为的影响。测定19种不同n端氨基酸残基α -氨基的pK(a)值。

Journal of chromatography Pub Date : 1993-08-27 DOI: 10.1016/s0021-9673(99)87003-4

T J Sereda, C T Mant, A M Quinn, R S Hodges

{"title":"Effect of the alpha-amino group on peptide retention behaviour in reversed-phase chromatography. Determination of the pK(a) values of the alpha-amino group of 19 different N-terminal amino acid residues.","authors":"T J Sereda, C T Mant, A M Quinn, R S Hodges","doi":"10.1016/s0021-9673(99)87003-4","DOIUrl":"https://doi.org/10.1016/s0021-9673(99)87003-4","url":null,"abstract":"We have examined the contribution of the alpha-amino group to retention behaviour for peptides in reversed-phase chromatography using two series of peptide analogues, one containing an N alpha-acetylated terminal and the other containing an alpha-amino group (non-acetylated). The effect of the alpha-amino group, at pH 2, on the hydrophobicity of the side-chain of the N-terminal residue was obtained by referencing the retention time of the acetylated or non-acetylated peptide to the retention time of a glycine analogue. It was shown that the presence of an alpha-amino group could decrease or increase the hydrophobicity of the side-chain of the N-terminal residue with respect to the hydrophobicity of the side-chain in the absence of an alpha-amino group. The effect was also shown to be sequence dependent, with respect to the N-terminal residue. Increasing pH was shown to increase retention time dramatically for the non-acetylated analogues, through the deprotonation of the alpha-amino group. By separating pairs of acetylated/non-acetylated analogues over the pH range 2-9, it was possible to determine the pK(a) of the alpha-amino group, where it was shown that the pK(a) was dependent on two probable factors: (1) the inherent hydrophobicity of the stationary phase; and (2) the amino acid substituted in the N-terminal position. Interestingly, the pK(a) values determined were very similar to that found in proteins. It was also possible to determine the pK(a) values of some of the substituted amino acids containing ionizable side-chains. This study shows that, in order to understand fully the retention behaviour of peptides containing an alpha-amino group in reversed-phase chromatography, one must incorporate an alpha-amino group contribution and its effect on the hydrophobicity of the side-chain of the N-terminal residue.","PeriodicalId":15508,"journal":{"name":"Journal of chromatography","volume":"646 1","pages":"17-30"},"PeriodicalIF":0.0,"publicationDate":"1993-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/s0021-9673(99)87003-4","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19394031","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 77

Characterisation of TNF-alpha-related peptides by high-performance liquid chromatography-mass spectrometry and high-performance liquid chromatography-tandem mass spectrometry. 高效液相色谱-质谱联用和高效液相色谱-串联质谱联用对tnf α相关肽的表征。

Journal of chromatography Pub Date : 1993-08-27 DOI: 10.1016/s0021-9673(99)87020-4

P F Alewood, A J Bailey, R I Brinkworth, D Fairlie, A Jones

{"title":"Characterisation of TNF-alpha-related peptides by high-performance liquid chromatography-mass spectrometry and high-performance liquid chromatography-tandem mass spectrometry.","authors":"P F Alewood, A J Bailey, R I Brinkworth, D Fairlie, A Jones","doi":"10.1016/s0021-9673(99)87020-4","DOIUrl":"https://doi.org/10.1016/s0021-9673(99)87020-4","url":null,"abstract":"Ion-spray triple quadrupole mass spectrometry and high-performance liquid chromatography were used to investigate the products from the solid phase synthesis of (H)-Leu-Thr-Glu-Asn-(OH), a TNF-alpha active-site probe. The target sequence was assembled using tert.-butoxycarbonyl (Boc) chemistry in stepwise fashion from the C-terminal on an Boc-Asn-OCH2-Pam-copoly(styrene-divinylbenzene) resin [Pam = 4-(carboxamidomethyl)benzyl ester]. The crude product was deprotected and cleaved from the resin by HF-p-cresol treatment for 1 h at 0 degrees C. HPLC analysis at 214 nm indicated two late-eluting major products and an early-eluting product. Preparative HPLC demonstrated that the early-eluting product contained ca. 80% of the expected recovered sample mass. Each component was then directly analysed by mass spectrometry and tandem mass spectrometry. The early eluting peak was confirmed as the desired LTEN sequence. Synthesis of the same sequence using 9-fluorenyl methoxycarbonyl (Fmoc) chemistry gave an identical product and confirmed the above analysis. The most significant by-product was derived from arylation of the glutamyl group by the quencher p-cresol. The likely origins of the by-products are discussed.","PeriodicalId":15508,"journal":{"name":"Journal of chromatography","volume":"646 1","pages":"185-91"},"PeriodicalIF":0.0,"publicationDate":"1993-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/s0021-9673(99)87020-4","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19394033","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 3

Human pepsin 3b peptide map sequence analysis, genotype and hydrophobic nature. 人胃蛋白酶3b肽图序列分析、基因型及疏水性。

Journal of chromatography Pub Date : 1993-08-27 DOI: 10.1016/s0021-9673(99)87022-8

A T Jones, J N Keen, N B Roberts

引用次数: 7

Analytical methods for differentiating minor sequence variations in related peptides. 鉴别相关肽序列微小变异的分析方法。

Journal of chromatography Pub Date : 1993-08-27 DOI: 10.1016/s0021-9673(99)87019-8

P A Grieve, A Jones, P F Alewood

{"title":"Analytical methods for differentiating minor sequence variations in related peptides.","authors":"P A Grieve, A Jones, P F Alewood","doi":"10.1016/s0021-9673(99)87019-8","DOIUrl":"https://doi.org/10.1016/s0021-9673(99)87019-8","url":null,"abstract":"A proline-rich peptide was isolated and purified to homogeneity from an extract of bovine neutrophil granules using semi-preparative RP-HPLC. The relative molecular mass of the peptide (called Bac-X) was determined by ionspray MS to be 5149 +/- 0.5. The amino acid composition of the peptide was characterized by its limited number of amino acid types, which included a high proline (43.3%) and arginine content (20.3%), and hydrophobic residues. Bac-X had similar characteristics to Bac-5, a previously characterised bactenecin of bovine neutrophil granules, with respect to its proline, arginine and hydrophobic amino acid content, molecular mass and antibacterial specificity. Tryptic and N-bromosuccinimide digestion of Bac-X produced fragments with masses (M(r) 785 and 4224 and 3100 respectively) consistent with those expected from a peptide with the reported sequence of Bac-5. Bac-X differed from Bac-5 in the number of amino acid residues (43 for Bac-X versus 42 for Bac-5) and contained glycine which Bac-5 did not. However, the calculated molecular mass of the peptide, based on the amino acid compositional data, did not match the experimental value. The purified peptide could not be sequenced by Edman degradation due to apparent blockage of the N-terminus. Partial sequence information, obtained by LC-MS and collision induced dissociation MS-MS analysis of a M(r) 785 tryptic fragment of Bac-X, showed that this peptide contained a six residue sequence (-RFPPIR-) not found in Bac-5 which, based on its reported sequence, contained a M(r) 785 tryptic fragment with the sequence -FRPPIR-. This difference in sequence of Bac-X compared with Bac-5 illustrates the application of electrospray (ionspray) MS techniques to the detection and identification of minor differences in related protein/peptide forms.","PeriodicalId":15508,"journal":{"name":"Journal of chromatography","volume":"646 1","pages":"175-84"},"PeriodicalIF":0.0,"publicationDate":"1993-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/s0021-9673(99)87019-8","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19394032","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 7

Structural characterisation of native and recombinant forms of the neurotrophic cytokine MK. 天然和重组形式的神经营养细胞因子MK的结构特征。

Journal of chromatography Pub Date : 1993-08-27 DOI: 10.1016/s0021-9673(99)87023-x

L Fabri, H Maruta, H Muramatsu, T Muramatsu, R J Simpson, A W Burgess, E C Nice

{"title":"Structural characterisation of native and recombinant forms of the neurotrophic cytokine MK.","authors":"L Fabri, H Maruta, H Muramatsu, T Muramatsu, R J Simpson, A W Burgess, E C Nice","doi":"10.1016/s0021-9673(99)87023-x","DOIUrl":"https://doi.org/10.1016/s0021-9673(99)87023-x","url":null,"abstract":"The retinoic acid (RA)-inducible midkine (MK) gene encodes a heparin-binding protein which can induce neurite outgrowth in cultured mammalian embryonic brain cells. This cytokine shares 65% amino acid sequence identity with another RA-inducible cytokine, pleiotropin (PTN). Both proteins contain 10 conserved cysteine residues, all of which appear to be disulphide linked. MK and PTN are also rich in lysine and arginine residues rendering them susceptible to proteolysis during purification, and making large-scale preparation of these molecules inherently difficult. Recombinant MK has been expressed as a fusion protein using a pGEX vector transfected into E. coli. To enable refolding of MK, the fusion protein was stored in solution at 4 degrees C for 14 days in the presence of dithiothreitol (DTT). Thrombin cleavage of the fusion protein, post storage, typically generated 5 mg of MK per litre of bacterial pellet. To establish the structural integrity of the recombinant product, we have analysed the refolding kinetics and compared the disulphide bond assignment of recombinant MK with that of native MK and native PTN. The synergistic use of micropreparative HPLC, to separate and recover in small eluant volumes enzymatically derived peptide fragments, with matrix assisted laser desorption mass spectrometry (MALD-MS) and N-terminal sequence analysis has allowed the unambiguous identification of the disulphide bonded fragments of native and recombinant MK. The disulphide bond assignment of MK is C12-C36, C20-C45, C27-C49, C59-C91 and C69-C101, and is equivalent to that of PTN.","PeriodicalId":15508,"journal":{"name":"Journal of chromatography","volume":"646 1","pages":"213-25"},"PeriodicalIF":0.0,"publicationDate":"1993-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/s0021-9673(99)87023-x","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19392098","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 67

Isolation of carboxyl-termini and blocked amino-termini of viral proteins by high-performance cation-exchange chromatography. 高效阳离子交换色谱法分离病毒蛋白的羧基端和阻断氨基端。

Journal of chromatography Pub Date : 1993-08-27 DOI: 10.1016/s0021-9673(99)87021-6

J J Gorman, B J Shiell

{"title":"Isolation of carboxyl-termini and blocked amino-termini of viral proteins by high-performance cation-exchange chromatography.","authors":"J J Gorman, B J Shiell","doi":"10.1016/s0021-9673(99)87021-6","DOIUrl":"https://doi.org/10.1016/s0021-9673(99)87021-6","url":null,"abstract":"The strong cation-exchanger, PolySulfoethyl Aspartamide, has been assessed as a medium for isolation of carboxyl-terminal and blocked amino-terminal peptides from tryptic digests of small quantities of viral proteins. Peptides with a single positive charge, the blocked amino-terminal peptides of ovalbumin and the Newcastle disease virus (NDV) matrix protein and carboxyl-terminal peptides of ovalbumin and the NDV nucleocapsid protein, eluted in early ion-exchange fractions and were readily isolated in homogeneous form by subsequent reversed-phase HPLC. Some early ion-exchange fractions also contained singly charged peptides derived by \"chymotryptic-like\" cleavage, whilst other peptides eluted in these fractions due to their highly acidic character. Terminal sequences with additional basic residues were isolated from later eluting ion-exchange fractions. Peptides with this property included the blocked amino-terminus of the NDV nucleocapsid protein and a portion of the carboxyl-terminus of the NDV matrix protein. Hitherto undescribed polymorphism in the amino-terminal region of ovalbumin was revealed in this study. Truncated peptides from the carboxyl-terminus of the NDV matrix protein were also detected. The presence of these peptides could be a reflection of carboxyl-terminal processing of the matrix protein. The strategy described herein should be of general utility for selective microisolation of carboxyl-terminal peptides and blocked amino-terminal peptides from tryptic digests of proteins.","PeriodicalId":15508,"journal":{"name":"Journal of chromatography","volume":"646 1","pages":"193-205"},"PeriodicalIF":0.0,"publicationDate":"1993-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/s0021-9673(99)87021-6","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19392096","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 19