Title : Chemical and functional characterization of a
fragment of
C1-s containing the epidermal growth
factor homology
region
Abstract :
- C1-s , one of the three subcomponents of C1-, the first component of complement, is a serine protease comprising two disulfide-linked chains, the B chain, containing the catalytic site , and the A chain, involved in Ca2 + binding and Ca2 (+)-dependent interaction(s) with the other C1- subcomponents
- In an attempt to identify the regions responsible for the latter functions, C1-s was submitted to limited proteolysis with plasmin , a treatment that split the A chain into three major fragments , alpha 1 , alpha 2, and gamma
- Fragment alpha 2, which comprised the epidermal growth factor-like ( EGF-like) region of C1-s , was heterogeneous, starting at serine 97 or phenylalanine 105 and ending at lysine 195
- This fragment was reduced and alkylated and then digested with elastase, and three peptides covering positions 131-135, 131-139, and 131-140 were characterized by amino acid analysis, Edman degradation, and mass spectrometry, showing that position 134 of C1-s is occupied partly by an asparagine (47 %) and partly by an erythro-beta-hydroxyasparagine, in contrast with the homologous position (150) of C1-r which only contains erythro-beta-hydroxyasparagine
- As measured by equilibrium dialysis, native alpha 2, like the other plasmin-cleavage fragments , did not retain the ability of intact C1-s to bind Ca2 +
- In the same way, plasmin cleavage abolished the ability of C1-s to dimerize or to associate with C1-r in the presence of Ca2 +
- In contrast, both alpha 2 and the N-terminal alpha 1 fragment , starting at serine 24 of the A chain, were able to compete significantly with intact C1s for the formation of the Ca2 (+)-dependent C1-s- C1r- C1-r- C1-s tetramer
- (ABSTRACT TRUNCATED AT 250 WORDS
Output (sent_index, trigger,
protein,
sugar,
site):
- 4. occupied, , -, -, position 134
Output(Part-Of) (sent_index,
protein,
site):
- 0. C1-s, fragment
- 0. C1-s, region
- 0. factor, region
- 3. C1-s, region
- 3. EGF, region
- 3. factor, region
- 4. C1-r, position
- 4. C1-s, position 134
- 5. plasmin, fragments
- 7. N-terminal alpha 1, fragment
*Output_Site_Fusion* (sent_index,
protein,
sugar,
site):