Molecular determinants of species specificity in the coronavirus receptor aminopeptidase N (CD13): influence of N-linked glycosylation.
Aminopeptidase N (APN), a 150-kDa metalloprotease also called CD13, serves as a receptor for serologically related coronaviruses of humans (human coronavirus 229E [HCoV-229E]), pigs, and cats. These virus-receptor interactions can be highly species specific; for example, the human coronavirus can use human APN (hAPN) but not porcine APN (pAPN) as its cellular receptor, and porcine coronaviruses can use pAPN but not hAPN. Substitution of pAPN amino acids 283 to 290 into hAPN for the corresponding amino acids 288 to 295 introduced an N-glycosylation sequon at amino acids 291 to 293 that blocked HCoV-229E receptor activity of hAPN. Substitution of two amino acids that inserted an N-glycosylation site at amino acid 291 also resulted in a mutant hAPN that lacked receptor activity because it failed to bind HCoV-229E. Single amino acid revertants that removed this sequon at amino acids 291 to 293 but had one or five pAPN amino acid substitution(s) in this region all regained HCoV-229E binding and receptor activities. To determine if other N-linked glycosylation differences between hAPN, feline APN (fAPN), and pAPN account for receptor specificity of pig and cat coronaviruses, a mutant hAPN protein that, like fAPN and pAPN, lacked a glycosylation sequon at 818 to 820 was studied. This sequon is within the region that determines receptor activity for porcine and feline coronaviruses. Mutant hAPN lacking the sequon at amino acids 818 to 820 maintained HCoV-229E receptor activity but did not gain receptor activity for porcine or feline coronaviruses. Thus, certain differences in glycosylation between coronavirus receptors from different species are critical determinants in the species specificity of infection.
Wentworth
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Department of Microbiology, University of Colorado Health Sciences Center, Denver, Colorado 80262, USA. Dave.Wentworth@UCHSC.edu
Holmes
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D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D001665:Binding Sites; D018826:CD13 Antigens / Q000737:chemistry / Q000378:metabolism*; D002460:Cell Line; D017934:Coronavirus / Q000378:metabolism / Q000472:pathogenicity*; D028941:Coronavirus 229E, Human; D018352:Coronavirus Infections / Q000821:virology; D016765:Coronavirus, Feline / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D000085064:Receptors, Coronavirus; D011991:Receptors, Virus / Q000737:chemistry / Q000378:metabolism*; D016415:Sequence Alignment; D013045:Species Specificity; D013329:Structure-Activity Relationship; D014162:Transfection; D005760:Transmissible gastroenteritis virus / Q000378:metabolism; D014774:Virulence
D008562:Membrane Glycoproteins; D000085064:Receptors, Coronavirus; D011991:Receptors, Virus; D018826:CD13 Antigens