title: Prominence of beta 2-microglobulin, class I heavy chain conformation, and tapasin in the interactions of class I heavy chain with calreticulin and the transporter associated with antigen processing.
abstract: Newly synthesized class I heavy (H) chain/beta 2m heterodimers awaiting peptides in the endoplasmic reticulum are associated with the transporter associated with Ag processing (TAP). We present evidence here that calreticulin, but not calnexin, displays steady state association with class I/TAP complexes. To separate the ability of beta 2m to bind with TAP and calreticulin from that of H chain, we studied human cell lines that lack expression of beta 2m or H chain. Little if any H chain was detected in association with TAP and calreticulin in the beta 2m- cell line Daudi. By contrast, high levels of beta 2m are found with TAP and calreticulin in the H chain-deficient cell line LCL 721.221, even after preclearance of the trace amount of class IB protein expressed by this cell line. Thus, beta 2m appears to bind TAP in the absence of H chain, providing an elegant mechanism to retain beta 2m in the endoplasmic reticulum at the site of peptide loading. To investigate whether other molecules participate in the binding of beta 2m and H chain to TAP and calreticulin, we analyzed the deletion mutant cell line LCL 721.220, which lacks tapasin. In 721.220, TAP and calreticulin are not associated with each other. Also, in these cells, H chain/beta 2m are not associated with TAP, but H chain and a low level of beta 2m are associated with calreticulin. These results suggest that tapasin is an obligatory mediator of the assemblage of calreticulin/H chain/beta 2m with TAP.
authors.affiliation: Department of Genetics, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: J C
authors.initials: JC
authors.lastname: Solheim
authors.forename: M R
authors.initials: MR
authors.lastname: Harris
authors.forename: C S
authors.initials: CS
authors.lastname: Kindle
authors.forename: T H
authors.initials: TH
authors.lastname: Hansen
mesh_terms: D000071181:ATP Binding Cassette Transporter, Subfamily B, Member 2; D018528:ATP-Binding Cassette Transporters / Q000276:immunology / Q000378:metabolism*; D017951:Antigen Presentation; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D037281:Calnexin; D037282:Calreticulin; D002460:Cell Line; D004721:Endoplasmic Reticulum / Q000378:metabolism; D015395:Histocompatibility Antigens Class I / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D011487:Protein Conformation; D012261:Ribonucleoproteins / Q000378:metabolism*; D013329:Structure-Activity Relationship; D001613:beta 2-Microglobulin / Q000378:metabolism*
chemical_list: D000071181:ATP Binding Cassette Transporter, Subfamily B, Member 2; D018528:ATP-Binding Cassette Transporters; D002135:Calcium-Binding Proteins; D037282:Calreticulin; D015395:Histocompatibility Antigens Class I; D012261:Ribonucleoproteins; C073119:TAP1 protein, human; D001613:beta 2-Microglobulin; D037281:Calnexin