Association of small ankyrin 1 with the sarcoplasmic reticulum

Abstract

Small ankyrin 1, or sAnk1, is a small, alternatively spliced product of the erythroid ankyrin gene, ANK1, that is expressed in striated muscle and concentrated in the network sarcoplasmic reticulum (SR) surrounding the Z disks and M lines. We have characterized sAnk1 in muscle homogenates and SR vesicles, and have identified the region that targets it to the network SR. Selective extractions and partitioning into Triton X-114 show that sAnk1 behaves like the SR Ca-ATPase and so is an integral protein of the SR membrane. Mild proteolytic treatment of isolated SR vesicles indicates that sAnk1 is oriented with its hydrophilic, C-terminal sequence exposed to the solution, which is equivalent to the cytoplasmic face of the SR membrane in situ. SDS-PAGE in non-reducing gels suggests that sAnk1 is present as dimers and larger oligomers in the native SR. These results suggest that sAnk1 is oligomeric and oriented with its C-terminus exposed to the cytoplasm, where it may interact with proteins of the contractile apparatus. The N-terminal 29 amino acid hydrophobic sequence of sAnk1, which is predicted to span the SR membrane, is sufficient to target proteins to and anchor them in internal membranes of HEK 293 cells. It also targets reporter proteins to the network SR of skeletal myofibers and is thus the first example of a sequence that targets proteins to a particular compartment of the SR.

• Ankyrin
• muscle
• sarcoplasmic reticulum
• tail-anchored protein
• targeting

Abbreviations
sAnk1	=	small product of the ankyrin 1 gene
SERCA	=	Ca-dependent ATPase of the sarcoplasmic reticulum
SR	=	sarcoplasmic reticulum
TCEP	=	tris (carboxyethyl) phosphine

Abbreviations
sAnk1	=	small product of the ankyrin 1 gene
SERCA	=	Ca-dependent ATPase of the sarcoplasmic reticulum
SR	=	sarcoplasmic reticulum
TCEP	=	tris (carboxyethyl) phosphine