TABLE 1.

KNRK and CHO fibroblast (Pro5 and Lec2) permanently expressing wild type (top) or N-linked glycosylation mutant (bottom) hPAR₁ cell clones generated for this study

In order to obtain permanent receptor-expressing cell lines, cells expressing high levels of hPAR₁ were isolated by FACS using the ATAP-2 anti-PAR₁ antibody. The glycosylation mutants were named with an N followed by a number related to the relative position of the potential glycosylation site and a Q. The alanine mutations were created at amino acid position Phe⁵⁵-Trp⁵⁶ or Val⁷²-Ser⁷³ of N62Q/N75Q to generate cathepsin G mutant (F55A/W56A) or proteinase 3 mutant (V72A/S73A).

Name of clone	Receptor cell surface expression level	Cell type
WT	Maximum level of WT hPAR1 cell surface expression	KNRK and CHO
WT hPAR1E	Maximum level of WT hPAR1E cell surface expression	KNRK and CHO
WT-H	Higher level of WT hPAR1 cell surface expression	KNRK
WT-M	Medium level of WT hPAR1 cell surface expression	KNRK
WT-L	Low level WT hPAR1 cell surface expression	KNRK

Name of clone	Consensus sequence(s) disrupted	Cell type
N35Q	Asn35	KNRK
N62Q	Asn62	KNRK
N75Q	Asn75	KNRK
N250Q	Asn250	KNRK
N259Q	Asn259	KNRK
N62Q/N75Q	Asn62 and Asn75	KNRK
N35Q/N62Q/N75Q	Asn35, Asn62, and Asn75	KNRK
N250Q/N259Q	Asn250 and Asn259	KNRK
N35–259Q	Asn35, Asn62, Asn75, Asn250, and Asn259	KNRK
N35QhPAR1E	Asn35	KNRK
N62Q/N75QhPAR1E	Asn62 and Asn75	KNRK
N250Q/N259QhPAR1E	Asn250 and Asn259	KNRK
N35–259QhPAR1E	Asn35, Asn62, Asn75, Asn250, and Asn259	KNRK
Cathepsin G mutant	Asn62, Asn75, Phe55, and Trp56	KNRK
Proteinase 3 mutant	Asn62, Asn75, Val72, and Ser73	KNRK