Sites on the cytoplasmic region of phospholamban involved in interaction with the calcium-activated ATPase of the sarcoplasmic reticulum.
Proton NMR studies have shown that when a peptide corresponding to the N-terminal region of phospholamban, PLB(1-20), interacts with the Ca2+ATPase of the sarcoplasmic reticulum, SERCA1a, docking involves the whole length of the peptide. Phosphorylation of Ser16 reduced the affinity of the peptide for the pump by predominantly affecting the interaction with the C-terminal residues of PLB(1-20). In the phosphorylated peptide weakened interaction occurs with residues at the N-terminus of PLB(1-20). PLB(1-20) is shown to interact with a peptide corresponding to residues 378-405 located in the cytoplasmic region of SERCA2a and related isoforms. This interaction involves the C-terminal regions of both peptides and corresponds to that affected by phosphorylation. The data provide direct structural evidence for complex formation involving residues 1-20 of PLB. They also suggest that phospholamban residues 1-20 straddle separate segments of the cytoplasmic domain of SERCA with the N-terminus of PLB associated with a region other than that corresponding to SERCA2a(378-405).
Levine
B A
BA
School of Biochemistry, University of Birmingham, UK. b.a.levine@bham.ac.uk
Patchell
V B
VB
Sharma
P
P
Gao
Y
Y
Bigelow
D J
DJ
Yao
Q
Q
Goh
S
S
Colyer
J
J
Drago
G A
GA
Perry
S V
SV
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002135:Calcium-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000252:Calcium-Transporting ATPases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D066298:In Vitro Techniques; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000201:enzymology*; D000099211:Phospholamban
D002135:Calcium-Binding Proteins; D000252:Calcium-Transporting ATPases; D010446:Peptide Fragments; D000099211:Phospholamban