Human neurotrophin-3: A one-step peptide mapping method and complete disulfide characterization of the recombinant protein

@article{Hui1996HumanNA,
  title={Human neurotrophin-3: A one-step peptide mapping method and complete disulfide characterization of the recombinant protein},
  author={John O Hui and John Le and V. Katta and Rakefet Rosenfeld and Michael F. Rohde and Mitsuru Haniu},
  journal={Journal of Protein Chemistry},
  year={1996},
  volume={15},
  pages={351-358}
}
Human neurotrophin-3 (NT-3) is a member of the nerve growth factor (NGF) family of neurotrophic factors, and the recombinant protein is being developed as a therapeutic for neurodegenerative diseases. The final product purity and lot-to-lot variation are monitored routinely by peptide mapping. However, only the N-terminal region of NT-3 was susceptible to proteolysis under native conditions. Complete digestion required that the protein be chemically modified by reduction and S-alkylation prior… 
NEUROTROPHIN EXPRESSION IN AGED PERIPHERAL NEURONS AND TARGETS by
TLDR
Dramatic alterations in NGF precursor proteins and significant increases in N GF transcript, together with minimal changes in NT-3, indicate that altered NGF regulation, particularly the presence of proNGF, contributes to the degeneration of aged sympathetic neurons.

References

SHOWING 1-10 OF 13 REFERENCES
Characterisation of neurotrophin dimers and monomers.
TLDR
Using this assay, the biological activities of guanidine-hydrochloride-denatured neurotrophin monomers were found to be much lower than that of the dimers, and experiments with NT-3 monomers and NIH3T3 cells expressing trkC suggest that such monomers exist in solution in a conformation that prevents efficient interactions with neurotrophIn receptors.
Nerve growth factor: Structure/function relationships
TLDR
The conclusion from these studies is that receptor interactions involve broad surface regions, which may be composed of residues from both protomers in the dimer, and specific surface lysine residues participate in receptor contacts.
Physicochemical Characterization of Recombinant Human Nerve Growth Factor Produced in Insect Cells with a Baculovirus Vector
TLDR
Recombinant human nerve growth factor secreted by insect cells was purified by ion‐exchange and reversed‐phase chromatography to near homogeneity and is suitable for further evaluation in animal models as a therapeutic molecule for neurodegenerative diseases such as Alzheimer's disease.
Nerve growth factor from mouse submaxillary gland: amino acid sequence.
  • R. Angeletti, R. Bradshaw
  • Chemistry, Biology
    Proceedings of the National Academy of Sciences of the United States of America
  • 1971
TLDR
The complete amino-acid sequence of the 2.5S nerve growth factor from male-mouse submaxillary glands has been determined and the alignment of the three disulfide bonds, determined from a combination of peptic and thermolytic digestions, is I-IV, II-V, and III-VI.
Peptide map analysis of recombinant human granulocyte colony stimulating factor: elimination of methionine modification and nonspecific cleavages.
TLDR
It is found that mild reduction and alkylation conditions can prevent methionine modification, while protease digestion in the presence of urea at room temperature alleviates generation of peptides derived from incomplete digestion and nonspecific cleavage by endoproteinase Glu-C.
Direct assignment of disulfide bonds by Edman degradation of selected peptide fragments.
Disulfide linkages in peptides or proteins were analyzed by automated gas-phase Edman sequencing performed in minimized reducing agents. If cystine linkage was regulated at the same position in two
Recombinant human brain-derived neurotrophic factor (rHuBDNF). Disulfide structure and characterization of BDNF expressed in CHO cells.
TLDR
Three disulfide linkages of recombinant human brain-derived neurotrophic factor (BDNF) were determined by peptide sequence analysis and characterized by mass spectrometry and the disulfides structure was homologous to that of nerve growth factor.
Identification of disulfide-bridged substructures within human von Willebrand factor.
TLDR
In the course of identifying substructural domains within the homooligomeric protein von Willebrand factor, seven large fragments of 8-90 kDa have been generated by limited proteolysis and it is shown that at least 35 disulfides must form intrachain bridges, specifically the cystines among residues 1-272 and 906-1492.
...
...