High-performance tryptic mapping of recombinant bovine somatotropin

Dougherty, John J.; Snyder, L. M.; Sinclair, Robert; Robins, Russell H.

doi:10.1016/0003-2697(90)90126-t

Cited by 27 publications

(16 citation statements)

References 9 publications

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“…Prior to tryptic digestion, the apolipophorins were denatured essentially as described by Matsudaira (1989) and Dougherty et al (1990). Briefly, 2 mg of the sample was concentrated in an Amicon cell (YM10 membrane), dialyzed against 0.05% sodium dodecyl sulfate (SDS) in 5 mM NH 4 HCO 3 at 4°C for 16 hours, and then precipitated with ice cold acetone, 1 mM HCl to remove excess SDS.…”

Section: Materials and Methods Polypeptide Isolation Tryptic Digestimentioning

confidence: 99%

Molecular characterization and gene expression in the eye of the apolipophorin II/I precursor fromLocusta migratoria

Bogerd

Babin

et al. 2000

J. Comp. Neurol.

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The transport of lipids via the circulatory system of animals constitutes a vital function that uses highly specialized lipoprotein complexes. In insects, a single lipoprotein, lipophorin, serves as a reusable shuttle for the transport of lipids between tissues. We have found that the two nonexchangeable apolipoproteins of lipophorin arise from a common precursor protein, apolipophorin II/I (apoLp-II/I). To examine the mechanisms of transport of lipids and liposoluble substances inside the central nervous system, this report provides the molecular cloning of a cDNA encoding the locust apoLp-II/I. We have recently shown that this precursor protein belongs to a superfamily of large lipid transfer proteins (Babin et al. [1999] J. Mol. Evol. 49:150-160). We determined that, in addition to its expression in the fat body, the locust apoLp-II/I is also expressed in the brain. Part of the signal resulted from fat body tissue associated with the brain; however, apoLp-II/I was strongly expressed and the corresponding protein detected, in pigmented glial cells of the lamina underlying the locust retina and in cells or cellular processes interspersed in the basement membrane. The latter finding strongly suggests an implication of apolipophorins in the transport of retinoids and/or fatty acids to the insect retina.

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Section: Materials and Methods Polypeptide Isolation Tryptic Digestimentioning

confidence: 99%

Molecular characterization and gene expression in the eye of the apolipophorin II/I precursor fromLocusta migratoria

Bogerd

Babin

et al. 2000

J. Comp. Neurol.

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“…In addition, the fragment T 10 co-eluted with fragment T 24 and fragment T 12 co-eluted with incompletely digested peptide T 15 -T 16 The presence of incompletely digested tryptic fragments in growth hormone digestion has been reported in the literature. [3][4][5] Some incompletely digested fragments (T 1 -T 2 , T 3 -T 4 , T 12 -T 13 , T 15 -T 16 , T 16 -T 17 , and T 20 -T 21 ) were observed in the tryptic peptide mapping of somidobove. In order to improve the digestion of some combined fragments, the incubation time in the digestion was increased to 15 hours at room temperature.…”

Section: Resultsmentioning

confidence: 96%

“…A technique using the combination of high-performance displacement chromatography with continuous-flow fast-atom bombardment (FAB) mass spectrometry has been used for the characterization of tryptic peptides of recombinant human growth hormone, 4 but this method is difficult to use in routine analysis of proteins. A thermospray ionization source mass spectrometry coupled with RPHPLC was used for the characterization of recombinant bovine growth hormone, 5 but this method met with limited success due to the thermal decomposition of peptides. Electrospray ionization mass spectrometry (ESI-MS) has rapidly matured into a powerful detection method for liquid chromatography of proteins and peptides.…”

mentioning

confidence: 99%

Separation and characterization of the tryptic peptide mapping of recombinant bovine growth hormone by reversed-phase high-performance liquid chromatography electrospray mass spectrometry

Chang

Kiehl

Kennington

1997

Rapid Commun. Mass Spectrom.

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Reversed phase high-performance liquid chromatography (RPHPLC) coupled with electrospray ionization mass spectrometry (ESI-MS) was used to separate and characterize the peptides resulting from the tryptic cleavage of somidobove, a recombinant bovine growth hormone. The tryptic digestion of somidobove was carried out at room temperature for 15 hours. The tryptic peptides were separated on a Zorbax SB300 C8 column with trifluoroacetic acid--acetonitrile gradient elution and characterized by LC/ESI-MS. Resulting single and combined peptide fragments were characterized by comparing the observed molecular mass with the calculated mass for these fragments.

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“…The digestion of control and oxidized bGH was performed according to the method optimized by Dougherty et al (15). A trypsin:rbGH ratio of 1:30 was adjusted for oxidized and control samples.…”

Section: Methodsmentioning

confidence: 99%

Characterization of the Metal-Binding Site of Bovine Growth Hormone through Site-Specific Metal-Catalyzed Oxidation and High-Performance Liquid Chromatography–Tandem Mass Spectrometry

Hovorka¹,

Williams²,

Schöneich³

2002

Analytical Biochemistry

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High-performance tryptic mapping of recombinant bovine somatotropin

Cited by 27 publications

References 9 publications

Molecular characterization and gene expression in the eye of the apolipophorin II/I precursor fromLocusta migratoria

Molecular characterization and gene expression in the eye of the apolipophorin II/I precursor fromLocusta migratoria

Separation and characterization of the tryptic peptide mapping of recombinant bovine growth hormone by reversed-phase high-performance liquid chromatography electrospray mass spectrometry

Characterization of the Metal-Binding Site of Bovine Growth Hormone through Site-Specific Metal-Catalyzed Oxidation and High-Performance Liquid Chromatography–Tandem Mass Spectrometry

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