Human Keratinocyte Growth Factor Recombinantly Expressed in Chinese Hamster Ovary Cells: Isolation of Isoforms and Characterization of Post-Translational Modifications

Hsu, Yueh-Rong; Hsu, Eric; Katta, Viswanatham; Brankow, David; Tseng, Julia; Hu, Sylvia; Morris, Charles F.; Kenney, William C.; Lu, Hsieng S.

doi:10.1006/prep.1997.0840

Cited by 30 publications

(24 citation statements)

References 27 publications

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“…Interestingly, ligands of the FGF7 subfamily contain an Arg-Xaa-Xaa-Arg cleavage motif in the N-terminal region preceding the β-trefoil core domain, and cultured mammalian cells secrete both full-length FGF7 ligand and the C-terminal fragment that results from proteolytic cleavage at the Arg-Xaa-Xaa-Arg motif 64 . However, whether specific proteolytic cleavage of FGF7 subfamily ligands occurs in vivo and is crucial for regulating the activity of these ligands is currently unknown.…”

Section: Regulation Of Fgf Ligand Activitymentioning

confidence: 99%

Exploring mechanisms of FGF signalling through the lens of structural biology

Goetz

Mohammadi

2013

Nat Rev Mol Cell Biol

486

464

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Fibroblast growth factors (FGFs) mediate a broad range of functions in both the developing and adult organism. The accumulated wealth of structural information on the FGF signalling pathway has begun to unveil the underlying molecular mechanisms that modulate this system to generate a myriad of distinct biological outputs in development, tissue homeostasis and metabolism. At the ligand and receptor level, these mechanisms include alternative splicing of the ligand (FGF8 subfamily) and the receptor (FGFR1–FGFR3), ligand homodimerization (FGF9 subfamily), site-specific proteolytic cleavage of the ligand (FGF23), and interaction of the ligand and the receptor with heparan sulphate cofactor and Klotho co-receptor.

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Section: Regulation Of Fgf Ligand Activitymentioning

confidence: 99%

Exploring mechanisms of FGF signalling through the lens of structural biology

Goetz

Mohammadi

2013

Nat Rev Mol Cell Biol

486

464

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“…1a and b. The spot trains in 2-D PAGE have been explained by posttranslational modifications like glycosylation [12], phosphorylation [13], carbamylation, truncation, or processing of several amino acid residues and many other modifications.…”

Section: Introductionmentioning

confidence: 99%

Deamidation as a widespread phenomenon in two-dimensional polyacrylamide gel electrophoresis of human blood plasma proteins

et al. 2000

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The human plasma protein patterns obtained by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) is a good model system for post-translational modifications because of the existence of several "ladders" of protein spots [Anderson, N. L., Anderson, N. G., Electrophoresis 1991, 12, 883-906], so-called "trains" of spots. Our investigation of several proteins, amongst others beta2-microglobulin and the haptoglobin chains, found the differences in isoelectric points (p/) to be due to deamidation of asparagines. After enzymatic cleavage with endopeptidases in the 2-D polyacrylamide gel, the asparagine and deamidated asparagine containing peptides were separated and quantified by reversed-phase HPLC. In order to separate these peptides, a neutral pH system was established and, as a result, the differences in hydrophobicity of asparagine-containing and deamidated asparagine-containing peptides increased. But how do deamidated asparagines contribute to the observed spot pattern? One spot in the 2-D gel consists of a mixture of protein species with the same number of deamidated asparagines but on different sequence position sites. The difference between the spots in the "ladder" is a growing number of negative charges introduced in the protein by an increasing number of deamidated asparagines. As a consequence, the mass difference between two spots is exactly 1 Da, which is shown in this paper for intact protein masses and the corresponding deamidated peptides.

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“…In addition to heterogeneous N-glycosylation, several posttranslational modifications, such as partial oxidation, partial sulfation, 0-glycosylation were known in KGF recombinantly expressed in Chinese hamster ovary cells [9]. The broad bands shown in this study may reflect such posttranslational modifications.…”

Section: Resultsmentioning

confidence: 60%

Evidence for the Presence Keratinocyte Growth Factor(KGF) in Human Ovarian Follicles.

et al. 2001

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Abstract.The presence of keratinocyte growth factor (KGF) in human follicular fluid (FF) was investigated in a total of 145 FFs obtained during oocyte retrieval for in vitro fertilization (IVF) from 29 patients with no apparent endocrine disorders.The concentrations of KGF, estradiol, progesterone, testosterone and human chorionic gonadotropin (hCG) in FF were measured by enzyme-linked immunosorbent assay. FF samples contained relatively higher amounts of KGF (2194± 87 pg/ml), whereas its concentrations in serum were below assay limit (< 31.2 pg/ml). Concentrations of KGF in FF were positively correlated with both progesterone (r=0.311, p<0.0005) and testosterone (r = 0.230, p < 0.01) concentrations in FF. However, KGF concentrations were not significantly correlated with estradiol and hCG concentrations. KGF in FF was detected as a broad band (26-29 kD) by immunoblotting, the size being reduced by 7kD after N-glycosidase treatment.In an in vitro experiment, KGF suppressed the basal and hCG-stimulated progesterone production by cultured human luteinized granulosa cells. In summary, we demonstrated the presence of KGF in human ovarian follicles, suggesting its possible role as a local factor in regulating human ovarian functions.

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Human Keratinocyte Growth Factor Recombinantly Expressed in Chinese Hamster Ovary Cells: Isolation of Isoforms and Characterization of Post-Translational Modifications

Cited by 30 publications

References 27 publications

Exploring mechanisms of FGF signalling through the lens of structural biology

Exploring mechanisms of FGF signalling through the lens of structural biology

Deamidation as a widespread phenomenon in two-dimensional polyacrylamide gel electrophoresis of human blood plasma proteins

Evidence for the Presence Keratinocyte Growth Factor(KGF) in Human Ovarian Follicles.

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