Characterization of human angiogenin variants implicated in amyotrophic lateral sclerosis.
Human angiogenin (ANG), the first member of the angiogenin family (from the pancreatic ribonuclease A superfamily) to be identified, is an angiogenic factor that induces neovascularization. It has received much attention due to its involvement in the growth of tumors and its elevated expression level in pancreatic and several other cancers. Recently the biological role of ANG has been shown to extend to the nervous system. Mutations in ANG have been linked with familial as well as sporadic forms of amyotrophic lateral sclerosis (ALS), a fatal neurodegenerative disorder characterized by selective destruction of motor neurons. Furthermore, mouse angiogenin-1 has been shown to be expressed in the developing nervous system and during the neuronal differentiation of pluripotent stem cells. We have now characterized the seven variants of ANG reported in ALS patients with respect to the known biochemical properties of ANG and further studied the biological properties of three of these variants. Our results show that the ribonucleolytic activity of six of the seven ANG-ALS implicated variants is significantly reduced or lost and some variants also show altered thermal stability. We report a significant reduction in the cell proliferative and angiogenic activities of the three variants that we chose to investigate further. Our studies on the biochemical and structural features of these ANG variants now form the basis for further investigations to determine their role(s) in ALS.
Crabtree
Benedict
B
Department of Biology and Biochemistry, University of Bath, Claverton Down, Bath BA2 7AY, United Kingdom.
Thiyagarajan
Nethaji
N
Prior
Stephen H
SH
Wilson
Peter
P
Iyer
Shalini
S
Ferns
Tyrone
T
Shapiro
Robert
R
Brew
Keith
K
Subramanian
Vasanta
V
Acharya
K Ravi
KR
D000690:Amyotrophic Lateral Sclerosis / Q000209:etiology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D004795:Enzyme Stability / Q000235:genetics; D005562:Formazans / Q000378:metabolism; D014644:Genetic Variation; D006801:Humans; D008545:Melanoma / Q000378:metabolism; D008958:Models, Molecular; D008970:Molecular Weight; D009154:Mutation; D011489:Protein Denaturation; D015202:Protein Engineering; D020673:Protein Renaturation; D012259:Ribonuclease, Pancreatic / Q000032:analysis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D013696:Temperature; D013778:Tetrazolium Salts / Q000378:metabolism
D005562:Formazans; D013778:Tetrazolium Salts; C079782:MTT formazan; C046366:angiogenin; D012259:Ribonuclease, Pancreatic