43 

 N-terminal sequencing 



About 1 pmol of purified SP-I protein was prepared for 

 N- terminal sequencing on SDS PAGE, followed by transfer of 

 the protein onto 3MM paper by electrophoresis. Mini-blot 

 system protocol and amino acid analysis by acid hydrolysis 

 was provided by the Protein Chemistry Core Facility, 

 Interdisciplinary Center of Biotechnology Research, 

 University of Florida. 

 Computer analysis 



The GCG computer software package (26) was used to 

 analyze nucleotide and deduced amino acid sequences of 

 cloned cDNAs . 



Biochemicals 

 Arogenate (90% pure) was isolated from a multiple 

 auxotroph of Neurospora crassa ATCC 36373 (42) or isolated 

 from a tyrosine auxotroph of Salmonella typhimurium (4) . 

 Prephenate (85% pure) was prepared as the barium salt from 

 culture supernatants of the tyrosine auxotroph of S. 

 typhimurium and was converted to the potassium salt with 

 excess K2SO4 prior to use. Dehydroquinate (90% pure) was 

 chemically synthesized by following the method of Haslam et 

 al . (37) using a platinum-catalyzed dehydrogenation of 

 quinic acid (about 90% pure) . 2' 5' ADP Sepharose-4B and AH- 

 Sepharose 4B were purchased from Pharmacia, Piscataway, NJ. 

 Bradford reagent was purchased from Bio-Rad, Rockville 



