Commercial recombinant human beta-nerve growth factor and adult rat dorsal root ganglia contain an identical molecular species of nerve growth factor prohormone.
Examination of commercial recombinant human beta-nerve growth factor (rh-beta-NGF) preparations with polyclonal antibodies specific to 13-kDa NGF and pro-NGF-specific domains revealed the presence of high-molecular-mass immunoreactive proteins, including a 60-kDa NGF prohormone. On incubation with a mixture of N- and O-specific glycosidases, the 60-kDa NGF pro-hormone generated a 32-kDa protein corresponding to the molecular size of NGF precursor predicted by the cloned human NGF cDNA. Highly sensitive chemiluminescence immunoblot analysis of adult rat dorsal root ganglia, spinal cord, and colon tissues with NGF- and pro-NGF domain-specific antibodies also revealed the presence of high-molecular-mass proteins, including the 60-kDa NGF prohormone. Based on the presence of the 60-kDa NGF prohormone in dorsal root ganglia and its efferent tissues, we suggest that proteolytically unprocessed, glycosylated NGF prohormone may mediate interactions between neurons and the tissues they innervate.
Reinshagen
M
M
Department of Medicine I, University of Ulm, Germany. max.reinshagen@medizin.uni-ulm.de
Geerling
I
I
Eysselein
V E
VE
Adler
G
G
Huff
K R
KR
Moore
G P
GP
Lakshmanan
J
J
D000818:Animals; D003106:Colon / Q000737:chemistry; D005727:Ganglia, Spinal / Q000737:chemistry*; D006801:Humans; D015151:Immunoblotting; D008163:Luminescent Measurements; D008970:Molecular Weight; D020932:Nerve Growth Factor / Q000032:analysis* / Q000737:chemistry*; D011498:Protein Precursors / Q000032:analysis* / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000737:chemistry; D013116:Spinal Cord / Q000737:chemistry
D011498:Protein Precursors; D011994:Recombinant Proteins; C061397:pro-nerve growth factor, human; D020932:Nerve Growth Factor