Self-assembly is important for TIP47 function in mannose 6-phosphate receptor transport

TY - JOUR

T1 - Self-assembly is important for TIP47 function in mannose 6-phosphate receptor transport

AU - Sincock, Paul M.

AU - Ganley, Ian G.

AU - Krise, Jeffrey P.

AU - Diederichs, Sven

AU - Sivars, Ulf

AU - O'Connor, Brian

AU - Ding, Li

AU - Pfeffer, Suzanne R.

PY - 2003/1/1

Y1 - 2003/1/1

N2 - TIP47 (tail-interacting protein of 47 kDa) binds to the cytoplasmic domains of mannose 6-phosphate receptors and is required for their transport from endosomes to the trans-Golgi network in vitro and in living cells. TIP47 occurs in cytosol as an oligomer; it chromatographs with an apparent mass of ∼300 kDa and displays an S-value of ∼13. Recombinant TIP47 forms homo-oligomers that are likely to represent hexamers, as determined by chemical cross-linking. Removal of TIP47 residues 1-151 yields a protein that behaves as a monomer upon gel filtration, yet is fully capable of binding mannose 6-phosphate receptor cytoplasmic domains. The presence of an oligomerization domain in the N-terminus of TIP47 was confirmed by expression of N-terminal residues 1-133 or 1-257 in mammalian cells. Co-expression of full-length TIP47 with either of these fragments led to the formation of higher-order aggregates of wild-type TIP47. Furthermore, the N-terminal domains expressed alone also occurred as oligomers. These studies reveal an N-terminal oligomerization domain in TIP47, and show that oligomerization is not required for TIP47 recognition of mannose 6-phosphate receptors. However, oligomerization is required for TIP47 stimulation of mannose 6-phosphate receptor transport from endosomes to the trans-Golgi in vivo.

AB - TIP47 (tail-interacting protein of 47 kDa) binds to the cytoplasmic domains of mannose 6-phosphate receptors and is required for their transport from endosomes to the trans-Golgi network in vitro and in living cells. TIP47 occurs in cytosol as an oligomer; it chromatographs with an apparent mass of ∼300 kDa and displays an S-value of ∼13. Recombinant TIP47 forms homo-oligomers that are likely to represent hexamers, as determined by chemical cross-linking. Removal of TIP47 residues 1-151 yields a protein that behaves as a monomer upon gel filtration, yet is fully capable of binding mannose 6-phosphate receptor cytoplasmic domains. The presence of an oligomerization domain in the N-terminus of TIP47 was confirmed by expression of N-terminal residues 1-133 or 1-257 in mammalian cells. Co-expression of full-length TIP47 with either of these fragments led to the formation of higher-order aggregates of wild-type TIP47. Furthermore, the N-terminal domains expressed alone also occurred as oligomers. These studies reveal an N-terminal oligomerization domain in TIP47, and show that oligomerization is not required for TIP47 recognition of mannose 6-phosphate receptors. However, oligomerization is required for TIP47 stimulation of mannose 6-phosphate receptor transport from endosomes to the trans-Golgi in vivo.

KW - Endosome

KW - Lysosome

KW - Mannose 6-phosphate receptor

KW - Membrane traffic

KW - TIP47

UR - https://www.scopus.com/pages/publications/0038445114

U2 - 10.1034/j.1600-0854.2003.40104.x

DO - 10.1034/j.1600-0854.2003.40104.x

M3 - Article

C2 - 12535272

AN - SCOPUS:0038445114

SN - 1398-9219

VL - 4

SP - 18

EP - 25

JO - Traffic

JF - Traffic

IS - 1

ER -