TABLE 2.

Primers for construction of gp78C mutants

To generate GST-gp78 mutants, human gp78C cDNA was amplified, and each mutant was subjected to site-directed mutagenesis with the following sense and antisense primers.

Constructs	Regions	Primers sequences
pGEX-gp78C	309–643 amino acids	5′-ggatcccgaattcgtcggcacaagaactat-3′
		5′-ctcgagggaggtctgctgcttctgaagcct-3′
pGEX-gp78-C1	309–574 amino acids	5′-ggatcccgaattcgtcggcacaagaactat-3′
		5′-ctcgagtctctcatcagcagacttggagaa-3′
pGEX-gp78-C2	309–452 amino acids	5′-ggatcccgaattcgtcggcacaagaactat-3′
		5′-ctcgagggagttgctggcctgcgtaatgcc-3′
pGEX-gp78-C3	309–343 amino acids	5′-ggatcccgaattcgtcggcacaagaactat-3′
		5′-ctcgagattgttgacagccagctcctctgg-3′
pGEX-gp78-K313A		5′-cgaattcgtcggcacgcgaactatctacgtg-3′
		5′-cacgtagatagttcgcgtgccgacgaattcg-3′
pGEX-gp78-K595A		5′-ctcctccagcaagctcgcgcacgtttcttgaacaaaag-′
		5′-ctttgttcaagaaacgtgcgcgagcttgctggaggag-3′
pGEX-gp78C-313PT	R307A/R308A	5′-gcgtggatccccaggagcagcgattcgtcggcaca-3′
		5′-tgtgccgacgaatcgctgctcctggggatccacgc-3′
	R310A/R311A	5′-gatccccaggaattgctgcgcacaagaactatcta-3′
		5′-tagatagttcttgtgcgcagcaattcctggggatc-3′
	H312A/K313A	5′-caggaattcgtcgggccgcgaactatctacgtgtg-3′
		5′-cacacgtagatagttcgcggcccgacgaattcctg-3′
pGEX-gp78C-595PT	R594A/K595A/R596A	5′-tccagcaagctggctgccgctttcttgaacaaaag-3′
		5′-cttttgttcaagaaagcggcagccagcttgctgga-3′
	R594V/K595A/R596V	5′-tccagcaagctggtcgccgtctttcttgaacaaaag-3′
		5′-cttttgttcaagaaagacggcgaccagcttgctgga-3′
pGEX-gp78C-586PT	Q584A/R585A/K586A	5′-gctggaggagttcgtccgcagccgccaccagcatgcgctgtc-3′
		5′-gacagcgcatgctggtggcggctgcggacgaactcctccagc-3′
pGEX-gp78C-ΔVIM	309–614 amino acids	5′-ggatcccgaattcgtcggcacaagaactat-3′
		5′-ctcgagtttaggggaggaagctctctgaggccgc-3′